JP2023086977A - Asymmetric synthesis of compounds and uses in disease treatments - Google Patents

Abstract

To provide methods of treating diseases.SOLUTION: The present application discloses, among other things, asymmetric synthesis of a diastereomeric compound of formula (I) (e.g., α-anordrin) or a salt thereof. Also provided are methods and compositions for treatment of estrogen deficiency and preventing or mitigating an estrogen deficiency symptom using a diastereomeric compound of formula (I) (e.g., α-anordrin) or a salt thereof alone or in combination with at least one additional agent. Further provided are methods and compositions for mitigating a side effect of an additional agent in the context of combination therapy with a diastereomeric compound of formula (I) (e.g., α-anordrin) or a salt thereof.SELECTED DRAWING: Figure 6

Description

TECHNICAL FIELD This invention relates to the field of asymmetric synthesis of steroid-like compounds, and more specifically to methods for preparing chiral compounds of anordrine and its analogues. The invention also relates to the therapeutic use of chiral compounds in the replacement treatment of estrogen deficiency symptoms.

BACKGROUND Decreased estrogen production in women on ovariectomy (OVX) or postmenopausal or anti-estrogen therapy leads to estrogen deficiency symptoms that can adversely affect quality of life for decades. Since the 1940's, estrogen replacement therapy (ERT) has been utilized to treat these conditions.

Estrogen binds to its receptors in many tissues during mammalian reproduction and development to regulate RNA transcription, stimulate cell proliferation, and modulate metabolic signaling. Three genes for estrogen binding proteins have been identified, which encode estrogen receptors (ER) α and β, and G protein-coupled estrogen receptor 1 (GPER1). ER-α and β have similar structural and functional domains, these domains being activation functional domain 1 (AF-1), DNA binding domain (DBD), dimerization domain and activation function. It contains domain 2 (AF-2), which is the ligand binding domain (LBD). They both belong to the nuclear superfamily of ligand-dependent transcription factors and have highly conserved DBD and LBD regions. They regulate RNA transcription upon ligand binding resulting in ligand-receptor complexes that can dimerize and translocate into the nucleus, where estrogens are found in the promoters of estrogen-responsive genes. Bind to response element (ERE). This type of modulation is typically referred to as the classical estrogen pathway. ER-α and β also regulate diverse biological functions through membrane-initiated estrogen signaling (MIES) and associate with the plasma membrane through interactions with their ligand-binding domains. The detailed molecular mechanisms of signaling by membrane-bound ER remain unclear. Modulatory effects of estrogen mediated by membrane-bound receptors on cell proliferation, matrix/migration, metabolism and glucose homeostasis have been reviewed (1, 2). Furthermore, studies on ER knockout mice have shown that ER-α is the predominantly functioning estrogen receptor compared to ER-β. Three transcriptional variants of ER-α (66, 46 and 36) have been found. ER-α36 lacks the AF-1 domain and contains a partial ligand-binding domain. It has been found to localize to the cell membrane and cytosol. ER-α-36 has limited ability to modulate MIES and was found to be uniquely expressed in tamoxifen-resistant cancer cells (such as MDA-MB-231 and Hec1A). MIES modulated by bound ER is thought to be responsible for the resistance to anti-estrogen therapy found by some investigators (3, 4).

However, studies showing that increased risk of breast and uterine cancer and the incidence of thromboembolism are associated with ERT have led to a decline in its use. Combined administration of estrogen and progesterone prevents breast and uterine cancer risk, but causes side effects of progesterone such as dizziness, nausea, vomiting, fatigue, anxiety, depression and headaches. Postmenopausal symptoms remain a problem for many older women. Therefore, there remains a continuing need for new developments of replacement treatments for estrogen deficiency symptoms.

SUMMARY OF THE INVENTION The present application, in one aspect, provides a method of synthesizing a diastereomeric compound of formula (I) (eg, α-anordrine) or a salt thereof. According to various embodiments described herein, the diastereomeric compound of formula (I) (eg, α-anordrine) or salt thereof is substantially pure.

In another aspect, a method of treating estrogen deficiency or preventing or alleviating estrogen deficiency symptoms, including OVX or postmenopausal symptoms, in an individual, wherein said individual comprises: a) a diastereomeric compound of formula (I); (e.g., alpha-anoldrin) or a salt thereof; and optionally b) an effective amount of at least one additional agent selected from the group consisting of selective estrogen receptor modulators and aromatase inhibitors. A method is provided comprising: In some embodiments, the additional agent is a selective estrogen receptor modulator selected from the group consisting of tamoxifen, raloxifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene ( SERMs). In some embodiments, the additional agent is an aromatase inhibitor selected from the group consisting of anastrozole, letrozole, exemestane, vorozole, formestane and fadrozole. In some embodiments, the estrogen deficiency symptom is selected from the group consisting of high liver triglycerides, osteoporosis, vulvovaginal atrophy, high blood triglycerides, high blood glucose and weight gain.

In some embodiments, a method of reducing the side effects of at least one additional agent with a diastereomeric compound of formula (I) (e.g., α-anoldrin) or a salt thereof, said method comprising: administering an effective amount of a diastereomeric compound of formula (I) (e.g., α-anoldrin) or a salt thereof in combination with an additional agent, wherein said additional agent is a selective estrogen receptor modulator and an aromatase inhibitor. A method is provided that is selected from the group of: In some embodiments, the additional agent is tamoxifen. In some embodiments, the additional agent is a selective estrogen receptor modulator selected from the group consisting of tamoxifen, raloxifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene ( SERMs). In some embodiments, the additional agent is an aromatase inhibitor selected from the group consisting of anastrozole, letrozole, exemestane, vorozole, formestane and fadrozole.

In some embodiments, the diastereomeric compound of Formula (I) or salt thereof and the additional agent are administered sequentially. In some embodiments, the diastereomeric compound of Formula (I) or salt thereof and the additional agent are co-administered.

In some embodiments, the individual is human.

In yet another aspect, a diastereomeric compound of formula (I) (e.g., α-anoldrin) or a salt thereof and at least one selected from the group consisting of selective estrogen receptor modulators (SERMs) and aromatase inhibitors. A pharmaceutical composition is provided that includes an additional agent. In some embodiments, the additional agent is tamoxifen. In some embodiments, the additional agent is raloxifene or a functional equivalent thereof (including, for example, raloxifene, lasofoxifene or bazedoxifene). In some embodiments, the additional agent is an aromatase inhibitor, such as anastrozole or a functional equivalent thereof.

In some embodiments, the weight ratio of the diastereomeric compound of formula (I) (eg, α-anoldrin) or salt thereof to the additional drug in the composition is from about 1:20 to about 20:1 (eg, , from about 10:1 to about 1:10 or from about 1:10 to about 1:15).

The pharmaceutical composition of the present invention may be in unit dosage form, such as oral unit dosage form, such as capsules, tablets, pills, caplets, gels, liquids (eg, suspensions, solutions, emulsions), powders or other particulate matter. can exist in

Also, use the pharmaceutical compositions described herein to treat estrogen deficiency or prevent or alleviate estrogen deficiency symptoms, including OVX or post-menopausal symptoms, as described herein. A method is provided.

These and other aspects and advantages of the present invention will become apparent from the detailed description below and the appended claims. It is to be understood that one, some or all features of the various embodiments described herein can be combined to form other embodiments of the invention.

Separation of α- and β-anordrin using silica gel. (FIG. 1A) Separation of α-anordrine (fraction 1 (F1)) and β-anordrine (fraction 2 (F2)) from the product of Synthetic Method I. (FIG. 1B) Only α-anordrine (Fraction 1 (F1)) is in the product of Synthetic Method II.

(FIG. 2A) α-Anordrin ((2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane) and (FIG. 2B) β-Anordrin ((2β,17α)- NMR analysis of diethynyl-(2α,17β)diol-dipropionate-A-nor-5α-androstane). (FIG. 2A) α-Anordrin ((2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane) and (FIG. 2B) β-Anordrin ((2β,17α)- NMR analysis of diethynyl-(2α,17β)diol-dipropionate-A-nor-5α-androstane).

Low dose α-anordrine (α-ANO) did not prevent uterine atrophy in OVX female mice compared to sham group. (FIG. 3A) Paraffin-embedded H&E section of mouse uterus treated with drug for 3 months (20× magnification). (Fig. 3B) Statistical analysis of wet weight of mouse uterus when measured from H&E stained sections as in (Fig. 3A). N=12. (FIGS. 3C, 3D and 3E) EEC mouse uterine height (μm) (shown in FIG. 3C), diameter ( mm) (shown in FIG. 3D) and circular muscle thickness (μm) (shown in FIG. 3E). N=12. Low dose α-anordrine (α-ANO) did not prevent uterine atrophy in OVX female mice compared to sham group. (FIG. 3A) Paraffin-embedded H&E section of mouse uterus treated with drug for 3 months (20× magnification). (Fig. 3B) Statistical analysis of wet weight of mouse uterus when measured from H&E stained sections as in (Fig. 3A). N=12. (FIGS. 3C, 3D and 3E) EEC mouse uterine height (μm) (shown in FIG. 3C), diameter ( mm) (shown in FIG. 3D) and circular muscle thickness (μm) (shown in FIG. 3E). N=12. Low dose α-anordrine (α-ANO) did not prevent uterine atrophy in OVX female mice compared to sham group. (FIG. 3A) Paraffin-embedded H&E section of mouse uterus treated with drug for 3 months (20× magnification). (Fig. 3B) Statistical analysis of wet weight of mouse uterus when measured from H&E stained sections as in (Fig. 3A). N=12. (FIGS. 3C, 3D and 3E) EEC mouse uterine height (μm) (shown in FIG. 3C), diameter ( mm) (shown in FIG. 3D) and circular muscle thickness (μm) (shown in FIG. 3E). N=12. Low dose α-anordrine (α-ANO) did not prevent uterine atrophy in OVX female mice compared to sham group. (FIG. 3A) Paraffin-embedded H&E section of mouse uterus treated with drug for 3 months (20× magnification). (Fig. 3B) Statistical analysis of wet weight of mouse uterus when measured from H&E stained sections as in (Fig. 3A). N=12. (FIGS. 3C, 3D and 3E) EEC mouse uterine height (μm) (shown in FIG. 3C), diameter ( mm) (shown in FIG. 3D) and circular muscle thickness (μm) (shown in FIG. 3E). N=12. Low dose α-anordrine (α-ANO) did not prevent uterine atrophy in OVX female mice compared to sham group. (FIG. 3A) Paraffin-embedded H&E section of mouse uterus treated with drug for 3 months (20× magnification). (Fig. 3B) Statistical analysis of wet weight of mouse uterus when measured from H&E stained sections as in (Fig. 3A). N=12. (FIGS. 3C, 3D and 3E) EEC mouse uterine height (μm) (shown in FIG. 3C), diameter ( mm) (shown in FIG. 3D) and circular muscle thickness (μm) (shown in FIG. 3E). N=12.

α-Anoldrin (α-ANO) exhibits dose-dependent activity in preventing vulvovaginal atrophy compared to β-anoldrin (β-ANO) in OVX female mice. (FIG. 4A) Paraffin-embedded H&E section of mouse vagina treated with drug for 3 months (20× magnification). (Fig. 4B and Fig. 4C) Wet weight (mg) of mouse uterus (shown in Fig. 4B) and vulvovaginal wall thickness (Fig. 4A), respectively, when measured from H&E-stained sections as in (Fig. 4A). μm) (shown in FIG. 4C). N=12. α-Anoldrin (α-ANO) exhibits dose-dependent activity in preventing vulvovaginal atrophy compared to β-anoldrin (β-ANO) in OVX female mice. (FIG. 4A) Paraffin-embedded H&E section of mouse vagina treated with drug for 3 months (20× magnification). (Fig. 4B and Fig. 4C) Wet weight (mg) of mouse uterus (shown in Fig. 4B) and vulvovaginal wall thickness (Fig. 4A), respectively, when measured from H&E-stained sections as in (Fig. 4A). μm) (shown in FIG. 4C). N=12. α-Anoldrin (α-ANO) exhibits dose-dependent activity in preventing vulvovaginal atrophy compared to β-anoldrin (β-ANO) in OVX female mice. (FIG. 4A) Paraffin-embedded H&E section of mouse vagina treated with drug for 3 months (20× magnification). (Fig. 4B and Fig. 4C) Wet weight (mg) of mouse uterus (shown in Fig. 4B) and vulvovaginal wall thickness (Fig. 4A), respectively, when measured from H&E-stained sections as in (Fig. 4A). μm) (shown in FIG. 4C). N=12.

α-Anoldrin exhibits dose-dependent activity in reducing blood glucose, TG levels and body weight compared to β-anoldrin in OVX female mice. (Fig. 5A) Statistical analysis of blood glucose (upper panel) and triglyceride (TG) (lower panel) levels. N=12. (Fig. 5B) Statistical analysis of mouse body weight (g) after drug administration. N=12. (Fig. 5C) Statistical analysis of initial body weight (g) of mice. N=12. NS means not significantly different. (Fig. 5D) Daily food intake by mice. N=12. NS means not significantly different. α-Anoldrin exhibits dose-dependent activity in reducing blood glucose, TG levels and body weight compared to β-anoldrin in OVX female mice. (Fig. 5A) Statistical analysis of blood glucose (upper panel) and triglyceride (TG) (lower panel) levels. N=12. (Fig. 5B) Statistical analysis of mouse body weight (g) after drug administration. N=12. (Fig. 5C) Statistical analysis of initial body weight (g) of mice. N=12. NS means not significantly different. (Fig. 5D) Daily food intake by mice. N=12. NS means not significantly different. α-Anoldrin exhibits dose-dependent activity in reducing blood glucose, TG levels and body weight compared to β-anoldrin in OVX female mice. (Fig. 5A) Statistical analysis of blood glucose (upper panel) and triglyceride (TG) (lower panel) levels. N=12. (Fig. 5B) Statistical analysis of mouse body weight (g) after drug administration. N=12. (Fig. 5C) Statistical analysis of initial body weight (g) of mice. N=12. NS means not significantly different. (Fig. 5D) Daily food intake by mice. N=12. NS means not significantly different. α-Anoldrin exhibits dose-dependent activity in reducing blood glucose, TG levels and body weight compared to β-anoldrin in OVX female mice. (Fig. 5A) Statistical analysis of blood glucose (upper panel) and triglyceride (TG) (lower panel) levels. N=12. (Fig. 5B) Statistical analysis of mouse body weight (g) after drug administration. N=12. (Fig. 5C) Statistical analysis of initial body weight (g) of mice. N=12. NS means not significantly different. (Fig. 5D) Daily food intake by mice. N=12. NS means not significantly different.

α-Anoldrin (α-ANO) exhibits dose-dependent activity in preventing TG accumulation in the liver compared to β-Anoldrin (β-ANO) in OVX female mice. (FIG. 6A) Paraffin-embedded H&E section of mouse liver treated with drug for 4 weeks (10× magnification). (FIG. 6B) Statistical analysis of TG content (mg/g) in mouse liver as measured from H&E-stained sections as in (FIG. 6A). N=12.

Micro-CT results show that α-anoldrin is more active in preventing osteoporosis compared to β-anoldrin in OVX female mice. (FIG. 7A) α-Anordrin shows higher activity in preventing loss of bone mean density (BMD) compared to β-Anordrin in OVX female mice. N=12. (FIG. 7B) α-Anordrin is more active in preventing a decrease in the ratio of bone volume (BV)/trabecular bone volume (TV) (BV/TV) compared to β-Anordrin in OVX female mice. indicates N=12.

α-Anoldrin inhibits tamoxifen-induced uterine EEC mitosis. TAM or a-ANO or b-ANO: Mice were treated with tamoxifen (TAM) or α-anordrine (a-ANO) or β-anordrine (b-ANO) alone. TAM+a-ANO or TAM+b-ANO mice were treated with a combination of tamoxifen and α-anordrine or β-anordrine, respectively. Blank mice were treated with vehicle. (FIG. 8A) Paraffin-embedded H&E section of EEC from mouse uterus treated with drug for 6 months (200× magnification). (FIG. 8B) Statistical analysis of EEC height (μm) when measured from H&E stained sections as in (A). N=6. ** means P<0.01. α-Anoldrin inhibits tamoxifen-induced uterine EEC mitosis. TAM or a-ANO or b-ANO: Mice were treated with tamoxifen (TAM) or α-anordrine (a-ANO) or β-anordrine (b-ANO) alone. TAM+a-ANO or TAM+b-ANO mice were treated with a combination of tamoxifen and α-anordrine or β-anordrine, respectively. Blank mice were treated with vehicle. (FIG. 8A) Paraffin-embedded H&E section of EEC from mouse uterus treated with drug for 6 months (200× magnification). (FIG. 8B) Statistical analysis of EEC height (μm) when measured from H&E stained sections as in (A). N=6. ** means P<0.01.

α-Anoldrine inhibits tamoxifen-induced NASH. TAM or a-ANO or b-ANO: Mice were treated with tamoxifen (TAM) or α-anordrine (a-ANO) or β-anordrine (b-ANO) alone. TAM+a-ANO or TAM+b-ANO mice were treated with a combination of tamoxifen and α-anordrine or β-anordrine, respectively. Blank mice were treated with vehicle. (FIG. 9A) Paraffin-embedded H&E section of drug-treated normal mouse liver (40× magnification). (FIG. 9B) Statistical analysis of liver triglyceride (TG) content as in (A); N=6; * and ** mean P<0.05 and P<0.01, respectively.

MicroCT results show that α-anordrine suppresses anastrozole-induced osteoporosis of bone. ANA: Mice were treated with Anastrozole (ANA) only. ANA + a-ANO or ANA + b-ANO: Mice were treated with a combination of anastrozole and an equal dose of α- or β-anordrine, respectively. Blank mice were treated with vehicle. (FIG. 10A) α-Anoldrine suppresses anastrozole-induced loss of bone mean density (BMD). N=6. ** indicates P<0.01. (FIG. 10B) α-Anordrin inhibits anastrozole-induced decrease in the ratio of bone volume (BV)/trabecular bone volume (TV) (BV/TV). N=6. ** indicates P<0.01. (FIG. 10C) α-Anordrin suppresses anastrozole-induced decrease in trabecular bone number (Tb.N). N=6. ** indicates P<0.01.

α-Anordrin suppresses anastrozole-induced vaginal atrophy. ANA: Mice were treated with Anastrozole (ANA) only. ANA + a-ANO or ANA + b-ANO: Mice were treated with a combination of anastrozole and an equal dose of α- or β-anordrine, respectively. Blank mice were treated with vehicle. (FIG. 11A) Paraffin-embedded H&E section of drug-treated normal mouse vagina (40× magnification). (FIGS. 11B and 11C) vulvovaginal wet weight (mg) (shown in FIG. 11B) and circular muscle thickness (μm) (shown in FIG. 11C), respectively. ) (FIG. 11A); N=6.

DETAILED DESCRIPTION OF THE INVENTION The present application provides methods for synthesizing diastereomeric compounds of formula (I) (eg, α-anordrine) or salts thereof. Also provided is the use of a diastereomeric compound of formula (I) (e.g., α-anordrin) or a salt thereof in treating estrogen deficiency in an individual, or preventing or alleviating estrogen deficiency symptoms, including OVX or postmenopausal symptoms. be. In addition, treatment of estrogen deficiency, reduction of side effects, or estrogen deficiency symptoms including OVX or postmenopausal symptoms including OVX or postmenopausal symptoms such as high liver triglycerides, osteoporosis, vulvovaginal atrophy, high blood triglycerides, high blood Compositions are provided for combination therapy comprising administering a diastereomeric compound of formula (I) (e.g., α-anordrine) or a salt thereof in conjunction with an additional agent for the prevention or reduction of glucose and weight gain. be done.

The present invention is based on the discovery of unique properties and mechanisms of action of diastereomeric compounds of formula (I) (eg, α-anoldrin) or salts thereof. In the present application, diastereomeric compounds of formula (I) (e.g. α-anoldrin) or salts thereof are compared to other diastereomeric compounds of formula (I) (e.g. β-anoldrin) or salts thereof , was recognized to be a more active and selective estrogen receptor modulator of membrane-bound estrogen-binding proteins. Beneficial effects of diastereomeric compounds of formula (I) (eg α-anoldrine) or salts thereof include i) estrogen deficiency symptoms including OVX or postmenopausal symptoms such as elevated liver triglycerides, osteoporosis, vulvovaginal atrophy, modulation of estrogen metabolic effects as agonists leading to prevention or reduction of high blood triglycerides, high blood glucose and weight gain; NASH), including organ atrophy and endometrial cancer).
definition

As those skilled in the art will recognize, two or more compounds that differ in stereochemistry only in the arrangement of their atoms in space are each considered diastereomers.

In one aspect, the compounds or salts thereof prepared by the methods of the present application can be substantially pure diastereomers. Compositions comprising the compounds detailed herein or salts thereof, eg, compositions comprising substantially pure diastereomeric compounds, are provided. By "substantially pure diastereomer" is intended a compound containing minor amounts of impurities, which means diastereomers other than the specified diastereomeric compound. In some embodiments, substantially pure diastereomeric compounds or salts thereof are provided in which the proportion of diastereomers or salts thereof is greater than or equal to about 98%. According to the present application, the percentage can be about 99%, about 99.5%, or about 99.9% or more.

It should be understood by those skilled in the art that the methods of combination therapy described herein require administration of one agent or composition in conjunction with the additional agent. "In conjunction with" means administering one treatment modality in addition to another treatment modality, e.g., administration of a second agent to the same individual in addition to a diastereomeric compound of formula (I) (e.g. , α-Anoldrin) or a salt thereof. Thus, "in conjunction with" refers to administration of one treatment modality prior to, during, or after delivery of the other treatment modality to the individual.

The methods described herein are generally useful for treating disease. As used herein, "treatment" is an approach for obtaining beneficial or desired clinical results. Alleviation of the pathological consequences of proliferative disorders is also included in "treatment". The methods of the invention contemplate any one or more of these aspects of treatment.

As used herein, the term "effective amount" is used to treat a particular disorder, condition or disease, e.g., ameliorate, alleviate, alleviate one or more of its symptoms, and/or It refers to an amount of a compound or composition sufficient to cause retardation. With respect to cancer, an effective amount is sufficient to cause tumor shrinkage and/or decrease the rate of tumor growth (e.g., slow tumor growth), or to prevent or retard other undesirable cell proliferation. Including quantity.

The term "individual" is a mammal, including humans. Individuals include, but are not limited to, humans, cows, horses, cats, dogs, rodents or primates. In some embodiments, the individual is human. In some embodiments the individual is an animal.

As used in this application, "alkyl" refers to a straight or branched chain saturated hydrocarbon. In some embodiments, alkyl groups have from 1 to 20 carbon atoms (“C ₁ -C ₂₀ alkyl”). In some embodiments, alkyl groups have from 1 to 12 carbon atoms (ie, (C ₁ -C ₁₂ alkyl)), or from 1 to 10 carbon atoms (ie, (C ₁ -C ₁₀ alkyl) ), or 1 to 8 carbon atoms (ie, (C ₁ -C ₈ alkyl)), or 1 to 6 carbon atoms (ie, (C ₁ -C ₆ alkyl)), or 1 to 4 It has carbon atoms (ie, (C ₁ -C ₄ alkyl)). Examples of suitable alkyl groups include, but are not limited to, methyl (Me, —CH ₃ ), ethyl (Et, —CH ₂ C
H ₃ ), 1-propyl (n-Pr, n-propyl, —CH ₂ CH ₂ CH ₃ ), 2-propyl (i-Pr, i-propyl, —CH(CH ₃ ) ₂ ), 1-butyl ( n-Bu, n-butyl, —CH ₂ CH ₂ CH ₂ CH ₃ ), 2-methyl-1-propyl (i-Bu, i-butyl, —CH ₂ CH(CH ₃ ) ₂ ), 2-butyl ( s-Bu, s-butyl, —CH(CH ₃ )CH ₂ CH ₃ ), 2-methyl-2-propyl (t-Bu, t-butyl, —C(CH ₃ ) ₃ ), 1-pentyl (n -pentyl, -CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2 -methyl-2-butyl (-C(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butyl ( —CH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butyl (—CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), 1-hexyl (—CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-hexyl (--CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (--CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-methyl- 2-pentyl (--C(CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl (--CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2- Pentyl (—CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl (—C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (— CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl (—C(CH ₃ ) ₂ CH(CH ₃ ) ₂ ), 3,3-dimethyl-2-butyl ( —CH(CH ₃ )C(CH ₃ ) ₃ and octyl (—(CH ₂ ) ₇ CH ₃ ).

"--Alkyl-" refers to a divalent radical derived from the above alkyl. In some embodiments, as used herein, -alkyl- is at least 1 carbon atom, at least 2 carbon atoms, at least 3 carbon atoms, at least 4 carbon atoms, at least 5 carbon atoms, at least 6 carbon atoms, at least 10 carbon atoms; at least 12 carbon atoms; at least 20 carbon atoms; or at least 40 carbon atoms; or 1-40 carbon atoms , 1 to 30 carbon atoms, 1 to 25 carbon atoms, 1 to 20 carbon atoms, 5 to 20 carbon atoms, 12 to 20 carbon atoms, or 14 to 18 carbon atoms . Examples of -alkyl- include, but are not limited to, methylene (-CH ₂ -), ethylene (-CH 2 CH ₂ -), propylene (-CH ₂ CH _{2 CH 2} -), butylene (-CH ₂ CH ₂ CH ₂ CH ₂ —).

As used herein, "alkenyl" refers to a straight or branched chain hydrocarbon having at least one carbon-carbon double bond. In some embodiments, an alkenyl group has 2 to 12 carbon atoms (ie, C ₂ -C ₁₂ alkenyl), or 2 to 10 carbon atoms (ie, C ₂ -C ₁₀ alkenyl), or 2 -8 carbon atoms (ie, C ₂ -C ₈ alkenyl), or 2-6 carbon atoms (ie, C ₂ -C ₆ alkenyl), or 2-4 carbon atoms (ie, C ₂ - _C4 alkenyl). Examples of suitable alkenyl groups include, but _are not limited to _, ethylene or vinyl (-CH= _CH2 ), allyl _{( -CH2CH} = _CH2 ) and 5-hexenyl ( _{-CH2CH2CH2CH2CH} = CH ₂ ).

"-Alkenyl-" refers to a divalent radical derived from the above alkenyl. In some embodiments, as used herein, -alkenyl- is at least 2 carbon atoms, at least 3 carbon atoms, at least 4 carbon atoms, at least 5 carbon atoms, at least 6 carbon atoms, at least 10 carbon atoms; at least 12 carbon atoms; at least 20 carbon atoms; or at least 40 carbon atoms; or 1-40 carbon atoms, 1-30 carbons atoms, 1 to 25 carbon atoms, 1 to 20 carbon atoms, 5 to 20 carbon atoms, 12 to 20 carbon atoms or 14 to 18 carbon atoms. By way of example, -alkenyl- can be -CH=CH-.

As used herein, "alkynyl" refers to an unsaturated straight or branched monovalent alkyl group having at least one site of acetylenic unsaturation (i.e., having at least one portion of the formula C≡C). Refers to a hydrocarbon chain or combination thereof. An alkynyl group can have a specified number of carbon atoms (ie, C ₂ -C ₂₀ means 2-20 carbon atoms). In some embodiments, an alkynyl group has 2-10 carbon atoms (ie, "C ₂ -C ₁₀ alkynyl") has 2-12 carbon atoms (ie, "C ₂ -C ₁₂ alkynyl") ”), has 2 to 8 carbon atoms (ie, “C ₂ -C ₈ alkynyl”), has 2 to 6 carbon atoms (ie, “C ₂ -C ₆ alkynyl”), or Those having 2 to 4 carbon atoms (ie, “C ₂ -C ₄ alkynyl”). Examples of alkynyl include, but are not limited to, ethynyl (or acetylenyl), prop-1-ynyl, prop-2-ynyl (or propargyl), but-1-ynyl, but-2-ynyl, but-3- Groups such as ynyl, homologs and isomers thereof, and the like are included.

The term "aryl" refers to and includes polyunsaturated aromatic hydrocarbon groups. Aryl can contain additional fused rings (eg, 1-3 rings), including further fused aryl, heteroaryl, cycloalkyl and/or heterocyclyl rings. In one variation, the aryl group contains 6-14 ring carbon atoms. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, biphenyl, and the like.

The term "cycloalkyl" can be fully saturated, monounsaturated or polyunsaturated but is non-aromatic and has the specified number of carbon atoms (eg, C ₁ -C ₁₀ has from 1 to 10 refers to and includes cyclic monovalent hydrocarbon structures having a Cycloalkyls can consist of a single ring, such as cyclohexyl, or multiple rings, such as adamantyl, but exclude aryl groups. Cycloalkyls containing more than one ring can be fused, spiro or bridged or combinations thereof. Preferred cycloalkyls are cyclic hydrocarbons having 3 to 13 ring carbon atoms. More preferred cycloalkyls are cyclic hydrocarbons having 3 to 8 ring carbon atoms (“C ₃ -C ₈ cycloalkyls”). Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, norbornyl, and the like.

"Halo" or "halogen" refers to Group 17 elements having atomic numbers 9-85. Preferred halo groups include fluoro, chloro, bromo and iodo. When a residue is substituted with more than one halogen, it may be designated by using a prefix corresponding to the number of attached halogen moieties. For example, dihaloaryl, dihaloalkyl, trihaloaryl, etc. are aryl and alkyl substituted with two (“di”) or three (“tri”) halo groups, which are not necessarily the same halo. thus 4-chloro-3-fluorophenyl is within the scope of dihaloaryl. Alkyl groups in which each hydrogen is replaced with a halo group are referred to as "perhaloalkyl." A preferred perhaloalkyl group is trifluoroalkyl (--CF ₃ ). Similarly, "perhaloalkoxy" refers to an alkoxy group in which each H in the hydrocarbon making up the alkyl portion of the alkoxy group is replaced by a halogen. An example of a perhaloalkoxy group is trifluoromethoxy (--OCF ₃ ).

The term "heteroaryl" refers to unsaturated aromatic cyclic groups having from 1 to 10 ring carbon atoms and at least one ring heteroatom including heteroatoms such as, but not limited to, nitrogen, oxygen and sulfur. refers to and includes, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule at a ring carbon or ring heteroatom. heteroaryl is further fused aryl,
It may contain additional fused rings (eg, 1-3 rings) including heteroaryl, cycloalkyl and/or heterocyclyl rings. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyrimidyl, thiophenyl, furanyl, thiazolyl, and the like.

The term "heterocycle" or "heterocyclyl" refers to a saturated or unsaturated non-aromatic group having 1 to 10 ring carbon atoms and 1 to 4 ring heteroatoms such as nitrogen, sulfur or oxygen. where the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized. Heterocyclyl groups can have a single ring or multiple condensed rings, but exclude heteroaryl groups. A heterocyclic ring containing more than one ring may be fused, spiro or bridged or any combination thereof. In fused ring systems, one or more of the fused rings can be aryl or heteroaryl. Examples of heterocyclyl groups include, but are not limited to, tetrahydropyranyl, dihydropyranyl, piperidinyl, piperazinyl, pyrrolidinyl, thiazolinyl, thiazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, 2,3-dihydrobenzo[b]thiophen-2-yl. , 4-amino-2-oxopyrimidin-1(2H)-yl and the like.

Unless otherwise specified, "optionally substituted" means that a group may be unsubstituted, and one or more of the substituents listed for that group (e.g., one, 2, 3, 4 or 5), and the substituents may be the same or different. In one embodiment, an optionally substituted group has one substituent. In another embodiment, an optionally substituted group has two substituents. In another embodiment, an optionally substituted group has 3 substituents. In another embodiment, an optionally substituted group has 4 substituents. In some embodiments, the optionally substituted groups are 1-2, 2-5, 3-5, 2-3, 2-4, 3-4, 1-3 1, 1 to 4 or 1 to 5 substituents.

"Pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation other than the active ingredient that is non-toxic to the subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers or preservatives.

Aspects and embodiments of the invention described herein are understood to include aspects and embodiments that "consist of" and/or "consist essentially of".

Reference to "about" a value or parameter herein includes (and describes) variations that are directed to that value or parameter per se. For example, reference to "about X" includes reference to "X."

As used in this specification and the appended claims, the singular forms "a,""or," and "the" include plural referents unless the context clearly dictates otherwise.
Compounds and Synthesis

The present disclosure includes specific stereochemical forms of the compounds described. In stereochemistry, each of two or more compounds that differ only in the arrangement of their atoms in space is considered a diastereomer. The diastereomeric compounds detailed herein can be substantially pure.

Generally speaking, chromatography, recrystallization and other conventional separation procedures may be employed with intermediates or final products in order to obtain specific isomers of a compound. In some embodiments, the diastereomeric compounds detailed herein can be prepared by resolution, in which the diastereomeric compound is separated from a mixture of diastereomers. In other embodiments,
Diastereomeric compounds are prepared by asymmetric syntheses that yield substantially pure diastereomeric compounds or salts thereof.

In some aspects, the present disclosure describes asymmetric syntheses that lead to substantially pure diastereomeric compounds or salts thereof. Provided herein are methods for preparing substantially pure diastereomeric forms of the compounds or salts thereof.

According to the present invention, the compounds are steroid-like compounds as detailed herein. For example, anoldrin is a steroid-like estrogen. In some embodiments, the diastereomeric compound is α-anordrine, (2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane).

（式中、
Ｒ_１は、－ＯＨ、－ＯＣ（Ｏ）－Ｒ^１ａまたは－ＯＣ（Ｏ）Ｒ^１ｂＣＯＯＨであり、Ｒ_４は、－ＯＨ、－ＯＣ（Ｏ）－Ｒ^４ａまたは－ＯＣ（Ｏ）Ｒ^４ｂＣＯＯＨであり、ここで、Ｒ^１ａおよびＲ^４ａは独立して、水素、Ｃ_１－Ｃ_６アルキル、Ｃ_２－Ｃ_６アルケニル、Ｃ_２－Ｃ_６アルキニルまたはＣ_３－Ｃ_６シクロアルキルであり、Ｒ^１ｂおよびＲ^４ｂは独立して、－Ｃ_１－Ｃ_６アルキル－または－Ｃ_２－Ｃ_６アルケニル－であり；
Ｒ_２およびＲ_５は、－Ｃ≡ＣＨであり；
Ｒ_６およびＲ_７は独立して、Ｃ_１－Ｃ_６アルキル、Ｃ_２－Ｃ_６アルケニル、Ｃ_２－Ｃ_６アルキニルまたはＣ_３－Ｃ_６シクロアルキルであり；
Ｒ_８およびＲ_９は独立して、水素、－ＯＨ、－ＮＨ_２、－ＮＯ_３、ハロゲン、Ｃ_１－Ｃ_６アルキル、Ｃ_２－Ｃ_６アルケニル、Ｃ_２－Ｃ_６アルキニル、Ｃ_３－Ｃ_６シクロアルキル、Ｃ_６－Ｃ_１４アリール、５～６員ヘテロアリールまたは３～６員ヘテロシクリルである）またはその塩を有する。 In some embodiments, the diastereomeric compound has the structure of Formula (I):

(In the formula,
R ₁ is -OH, -OC(O)-R ^1a or -OC(O)R ^1b COOH and R ₄ is -OH, -OC(O)-R ^4a or -OC(O)R ^4b COOH, wherein R ^1a and R ^4a are independently hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ cycloalkyl; R ^1b and R ^4b are independently —C ₁ -C ₆ alkyl- or —C ₂ -C ₆ alkenyl-;
R ₂ and R ₅ are —C≡CH;
R ₆ and R ₇ are independently C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ cycloalkyl;
R ₈ and R ₉ are independently hydrogen, —OH, —NH ₂ , —NO ₃ , halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₄ aryl, 5-6 membered heteroaryl or 3-6 membered heterocyclyl) or salts thereof.

In some embodiments, R ₁ is hydroxyl.

In some embodiments, R ₁ is -OC(O)R ^1a , where R ^1a is C ₁ -C ₆ alkyl. In some embodiments, R ^1a is C ₁ -C ₄ alkyl, such as methyl, ethyl, 1-propyl (n-Pr, n-propyl, —CH ₂ CH ₂ CH ₃ ), 2-propyl (i-Pr , i-propyl, —CH(CH ₃ ) ₂ ), 1-butyl. In some embodiments, R ^1a is C ₁ -C ₃ alkyl or C ₁ -C ₂ alkyl. In some embodiments, R ^1a is ethyl.

In some embodiments, R ₁ is -OC(O)R ^1b COOH, where R ^1b
is -C ₁ -C ₆ alkyl-. In some embodiments, R ₁ is -OC(O)R ^1b COOH, where R ^1b is -C ₂ -C ₆ alkenyl-. In some embodiments, R ₁ is -OC(O)R ^1b COOH, where R ^1b is methylene (ie, -CH ₂ -) and R ₁ is -OC(O)CH ₂ COOH is. As some examples of salt forms, in certain embodiments, R ₁ is -OC(O)CH ₂ COOLi, -OC(O)CH ₂ COONa, or -OC(O)CH ₂ COOK. In some embodiments, R ₁ is —OC(O)R ^1b COOH, where R ^1b is ethylene (ie, —CH ₂ CH ₂ —) and R ₁ is —OC(O)CH ₂ CH ₂ COOH. In certain embodiments, R ₁ is -OC(O)CH ₂ CH ₂ COOLi, -OC(O)CH ₂ CH ₂ COONa, or -OC(O)CH ₂ CH ₂ COOK. In some embodiments, R ₁ is -OC(O)R ^1b COOH, where R ^1b is -CH=CH- and R ₁ is -OC(O)CH=CHCOOH. In certain embodiments, R ₁ is -OC(O)CH=CHCOOLi, -OC(O)CH=CHCOONa or -OC(O)CH=CHCOOK.

In some embodiments, _R4 is hydroxyl.

In some embodiments, R ₄ is -OC(O)R ^4a , where R ^4a is C ₁ -C ₆ alkyl. In some embodiments, R ^4a is C ₁ -C ₄ alkyl, such as methyl, ethyl, 1-propyl (n-Pr, n-propyl, —CH ₂ CH ₂ CH ₃ ), 2-propyl (i-Pr , i-propyl, —CH(CH ₃ ) ₂ ), 1-butyl. In some embodiments, R ^4a is C ₁ -C ₃ alkyl or C ₁ -C ₂ alkyl. In some embodiments, R ^4a is ethyl.

In some embodiments, R ₄ is -OC(O)R ^4b COOH, wherein R ^4b is -C ₁ -C ₆ alkyl-. In some embodiments, R ₄ is -OC(O)R ^4b COOH, wherein R ^4b is -C ₂ -C ₆ alkenyl-. In some embodiments, R ₄ is -OC(O)R ^4b COOH, where R ^4b is methylene (ie, -CH ₂ -) and R ₄ is -OC(O)CH ₂ COOH is. As some examples of salt forms, in certain embodiments, R ₄ is -OC(O)CH ₂ COOLi, -OC(O)CH ₂ COONa, or -OC(O)CH ₂ COOK. In some embodiments, R ₄ is -OC(O)R ^4b COOH, where R ^4b is ethylene (ie, -CH ₂ CH ₂ -) and R ₁ is -OC(O)CH ₂ CH ₂ COOH. In certain embodiments, R ₄ is -OC(O)CH ₂ CH ₂ COOLi, -OC(O)CH ₂ CH ₂ COONa, or -OC(O)CH ₂ CH ₂ COOK. In some embodiments, R ₄ is -OC(O)R ^4b COOH, where R ^4b is -CH=CH- and R ₄ is -OC(O)CH=CHCOOH. In certain embodiments, R ₄ is -OC(O)CH=CHCOOLi, -OC(O)CH=CHCOONa or -OC(O)CH=CHCOOK.

In some embodiments, R ₆ is C ₁ -C ₆ alkyl. In some embodiments, R ₆ is C ₂ -C ₆ alkenyl. In some embodiments, R ₇ is C ₁ -C ₆ alkyl. In some embodiments, R ₇ is C ₂ -C ₆ alkenyl. R ₆ and R ₇ may or may not be the same.

In some embodiments, R ₈ is hydrogen, —OH, —NH ₂ , —NO ₃ , halogen, C ₁ -C ₆ alkyl or C ₂ -C ₆ alkenyl. In some embodiments, _R8 is hydrogen. In some embodiments, R ₈ is C ₁ -C ₆ alkyl. In some embodiments, R ₉ is hydrogen, —OH, —NH ₂ , —NO ₃ , halogen, C ₁ -C ₆ alkyl or C ₂ -C ₆ alkenyl. In some embodiments, _R9 is hydrogen. In some embodiments, R ₉ is C ₁ -C ₆ alkyl. R ₈ and R ₉ may or may not be the same.

（ここで、Ｒ_６、Ｒ_７、Ｒ_８およびＲ_９は、式（Ｉ）について定義されるとおりである）を式（ＩＩＩ）

のシリルアセチレンと反応させることを含む方法を使用して調製され得る。 A substantially pure diastereomeric compound or a salt thereof described herein can be prepared by diketone compound of formula (II) in the presence of an organometallic reagent R ₁₀ -M.

(wherein R ₆ , R ₇ , R ₈ and R ₉ are as defined for formula (I)) to formula (III)

can be prepared using a method comprising reacting with a silylacetylene of

R ₆ , R ₇ , R ₈ and R ₉ of the diketone compounds of formula (II) are as defined above, and in various embodiments of the compounds have the structure of formula (I).

With respect to Reaction Step III-2 of Method II shown in the Examples below, an exemplary process for preparing α-anoldrin is to react the diketone product (c) with a silylacetylene, trimethylsilylacetylene (TMS). including. The present application recognizes that silylacetylenes can be used for the asymmetric alkynylation of the deketone compounds described herein. Without being bound by any particular theory, it is believed that the two terminal alkynes add one each to the two carbonyl groups of the deketone compound.

R ^a , R ^b and R ^c of the silylacetylene of formula (III) can be the same or different. In some embodiments, R ^a , R ^b and R ^c are hydrogen, —OH, —OH, halogen or C ₁ -C ₂₀ alkyl optionally substituted with C ₁ -C ₆ alkyl, —OH , C ₁ -C ₆ alkoxy optionally substituted with halogen or C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₄ aryl, 5-6 membered heteroaryl or 3-6 membered independently selected from the group consisting of heterocyclyl; In some embodiments, R ^a , R ^b and R ^c are independently C ₁ -C ₂₀ alkyl optionally substituted with —OH, halogen or C ₁ -C ₆ alkyl. In some embodiments, R ^a , R ^b and R ^c are independently C ₁ -C ₆ alkyl, such as methyl, ethyl, 1-propyl (
n-Pr, n-propyl, —CH ₂ CH ₂ CH ₃ ), 2-propyl (i-Pr, i-propyl, —CH(CH ₃ ) ₂ ), 1-butyl. In some embodiments, R ^a , R ^b and R ^c are methyl, thus the silylacetylene is trimethylsilylacetylene (TMS). Without being bound by any particular theory, one or more of R ^a , R ^b and R ^c has a large size that creates steric hindrance, which converts one diastereomer to another It is thought that it causes to form more preferentially.

Organometallic reagents used according to this method are defined as R ₁₀ -M, where M is a metal. As an example, R ₁₀ -M is n-butyllithium.

In some embodiments, M is Li, Na or K. In some embodiments, M is Li.

In some embodiments, R ₁₀ is C ₁ -C ₂₀ alkyl optionally substituted with —OH, halogen or C ₁ -C ₆ alkyl. In some embodiments, R ₁₀ is C ₁ -C ₆ alkyl. In some embodiments, R ₁₀ is n-butyl.

In some embodiments, tetramethylethylenediamine (TEMED) is added with the organometallic reagent.

In various embodiments, the methods provided herein are performed in the presence of an organic solvent. A non-limiting list of suitable organic solvents includes, but is not limited to, dichloromethane, chloroform, acetonyltril, dichloroethane, tetrahydrofuran (THF), dimethylsulfoxide and toluene.

The methods provided herein are performed at relatively low reaction temperatures. The reaction temperature can range from about 0°C to about -100°C. In some embodiments, the reaction temperature is from about -20°C to about -90°C. In some embodiments, the reaction temperature is from about -40°C to about -80°C. In some embodiments, the reaction temperature is about -30°C to about -50°C.

を除去することをさらに含む。 According to some embodiments of the present invention, a method for stereospecifically preparing a substantially pure diastereomeric compound or a salt thereof comprises a silyl group of formula (IV):

further comprising removing

In some embodiments, the removing step is performed by contact with a deprotecting agent. A non-limiting list of suitable deprotecting agents includes, but is not limited to, tetrabutylammonium fluoride (TBAF), hydrofluoric acid and potassium fluoride.
Pharmaceutical compositions and formulations

The present disclosure includes specific diastereomeric forms of the compounds described herein. Also provided are salts, such as pharmaceutically acceptable salts, of the compounds referred to herein. All forms of the Diastereomeric Compounds are also included in the present invention, including crystalline or amorphous forms of the Diastereomeric Compounds. Compositions comprising diastereomeric compounds of the invention or salts thereof, such as compositions of substantially pure diastereomeric compounds or salts thereof, are also contemplated.

Pharmaceutical compositions of any of the diastereomeric compounds detailed herein are encompassed by the present disclosure. The present disclosure includes pharmaceutical compositions comprising substantially pure diastereomeric compounds of formula (I) or salts thereof and a pharmaceutically acceptable carrier or excipient. In one aspect, the salt is a pharmaceutically acceptable salt. Salts, for example, are acid addition salts, eg salts formed with inorganic or organic acids. Pharmaceutical compositions may take any form suitable for oral, buccal, parenteral, nasal, topical or rectal administration, or for administration by inhalation.

In one variation, a substantially pure diastereomeric compound or salt thereof is a synthetic product prepared for administration to an individual. In another variation, compositions containing substantially pure diastereomeric compounds or salts thereof are provided. In another variation, the disclosure includes pharmaceutical compositions comprising a compound detailed herein and a pharmaceutically acceptable carrier.

The diastereomeric compounds detailed herein or salts thereof can be delivered orally, mucosally (eg nasal, sublingual, vaginal, buccal or rectal), parenterally (eg intramuscularly, subcutaneously or intravenously). ) can be formulated for any available delivery route, including forms of delivery, topical delivery or transdermal delivery. Diastereomeric compounds or salts thereof include, but are not limited to, tablets, caplets, capsules (e.g., hard or soft gelatin capsules), cachets, troches, lozenges, gums, dispersions, suppositories, ointments, cataplasms (poultices), Pastes, powders, dressings, creams, solutions, patches, aerosols (e.g. nasal sprays or inhalers), gels, suspensions (e.g. aqueous or non-aqueous liquid suspensions, oil-in-water emulsions or water-in-oil emulsions) ), may be formulated with suitable carriers to provide delivery forms including solutions and elixirs.

The diastereomeric compounds or salts thereof described herein can be used in the preparation of formulations such as pharmaceutical formulations by combining the compounds or salts thereof as active ingredients with pharmaceutically acceptable carriers such as those described above. can be used. The carrier can be in a variety of forms, depending on the therapeutic form of the system (eg, transdermal patch versus oral tablet). In addition, the pharmaceutical formulation contains preservatives, solubilizers, stabilizers, rewetting agents, emulsifiers, sweeteners, dyes, modifiers and salts for adjusting osmotic pressure, buffers, coating agents or antioxidants. can. Formulations containing the compounds may also contain other substances with beneficial therapeutic properties. Pharmaceutical formulations may be prepared by known pharmaceutical methods. Suitable formulations are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, PA, ^20th ed. (2000), which is incorporated herein by reference.

In some embodiments, the diastereomeric compounds described herein are incorporated into generally acceptable oral compositions such as hard or soft shells, tablets in emulsions or suspensions, coated tablets and gel capsules. can be administered to an individual in the form Examples of carriers that may be used in preparing such compositions are lactose, corn starch or derivatives thereof, talc, stearate or salts thereof, and the like. Acceptable carriers for gel capsules with soft shells are, for example, vegetable oils, waxes, fats, semi-solid and liquid polyols, and the like. In addition, the pharmaceutical formulation contains preservatives, solubilizers, stabilizers, rewetting agents, emulsifiers, sweeteners, dyes, modifiers and salts for adjusting osmotic pressure, buffers, coating agents or antioxidants. can.

Any of the diastereomeric compounds described herein or salts thereof can be formulated in tablets of any dosage form described, e.g. Acceptable salts may be formulated as 10 mg tablets.
how to use

Compounds and compositions detailed herein, for example, diastereomeric compounds of formula (I) provided herein or salts thereof, and a pharmaceutically acceptable carrier or excipient Pharmaceutical compositions can be used in the methods of administration and treatment provided herein.

Estrogen deficiency is associated with the development of many health conditions such as infertility, premature aging, osteoporosis and cardiovascular disease. Ovariectomized (OVX) or post-menopausal individuals typically suffer from estrogen deficiency. Estrogen deficiency symptoms described herein include, but are not limited to, osteoporosis, fatty liver, weight gain, high blood triglycerides and blood glucose and organ atrophy (e.g. kidney, muscle and vulvovaginal atrophy). be done.

In some embodiments, a method of treating estrogen deficiency in an individual comprising administering to said individual an effective amount of a diastereomeric compound of formula (I) (e.g., α-anoldrin) or a salt thereof. A method is provided.

In some embodiments, a method of preventing or alleviating estrogen deficiency symptoms in an individual comprises administering to said individual an effective amount of a diastereomeric compound of formula (I) (e.g., alpha-anordrine) or a salt thereof. A method is provided comprising: In some embodiments, methods are provided for preventing or alleviating estrogen deficiency symptoms selected from the group consisting of high liver triglycerides, osteoporosis, vulvovaginal atrophy, high blood triglycerides, high blood glucose and weight gain.

Without being bound by any particular theory, it is observed that the diastereomeric compounds of formula (I) or salts thereof detailed herein possess therapeutic effects that other stereochemical forms do not possess. . Accordingly, the diastereomeric compounds of formula (I), compositions and methods thereof detailed in this application have advantageous therapeutic effects compared to other stereochemical forms or mixtures of compounds of formula (I). can have As one specific example, two chiral anordrin compounds named 2alpha-(α-) and 2beta-(β-) anordrin were found (18, 19). According to some embodiments of the present invention, α-anoldrin has therapeutic efficacy in treating estrogen deficiency and in preventing or alleviating estrogen deficiency symptoms, whereas β-anoldrin is minimally therapeutic in these treatments. has no effect.

In some embodiments, a diastereomeric compound of formula (I) (eg, α-anoldrin) or a salt thereof is co-administered with one or more additional agents, which can be (separately or simultaneously) Combination therapies are provided herein, for example, the diastereomeric compounds of formula (I) or salts thereof and one or more additional agents (e.g., pharmaceutical compositions described herein Additionally or alternatively, the diastereomeric compound of formula (I) or salt thereof and one or more additional agents may be administered sequentially, for example, 1 The one or more additional agents are administered first and the diastereomeric compound of formula (I) or salt thereof is administered next, or the diastereomeric compound of formula (I) or salt thereof is administered first administered, and one or more additional agents are then administered.

The present disclosure provides methods of treatment of estrogen deficiency, as well as methods of preventing or alleviating estrogen deficiency symptoms, comprising diastereomeric compounds of formula (I) (e.g., α-anordrine) in combination with at least one additional agent. or a method comprising administering a salt thereof. Additional agents may be selective estrogen receptor modulators (SERMs) and aromatase inhibitors.

SERMs are currently used as treatments for breast cancer, osteoporosis, and post-menopausal conditions, as these drugs have the property that they can act as estrogen agonists and antagonists, depending on the target tissue. According to the present invention, suitable SERMs include, but are not limited to tamoxifen, raloxifene, toremifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene. In some embodiments, the SERM used in combination with the diastereomeric compounds detailed herein or salts thereof is tamoxifen. In some embodiments, the SERM used in combination with a diastereomeric compound or salt thereof detailed herein is raloxifene or a functional equivalent thereof. As used herein, "functional equivalents thereof" refer to compounds that function through the same mechanism as raloxifene. For example, functional equivalents of raloxifene include, but are not limited to lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene.

In some embodiments, the method of treating estrogen deficiency comprises administering a diastereomeric compound of formula (I) (eg, α-anoldrine) or a salt thereof in combination with at least one SERM. In some embodiments, the method of preventing or alleviating estrogen deficiency symptoms comprises administering a diastereomeric compound (eg, α-anoldrine) or salt thereof in combination with at least one SERM.

"Aromatase inhibitors" refer to a class of drugs that inhibit aromatase activity. Aromatase inhibitors are used in the treatment of breast and ovarian cancer in postmenopausal women to reduce the increase in estrogen conversion during cycling with external testosterone. Suitable aromatase inhibitors include, but are not limited to, anastrozole (Arimidex), letrozole (Femara), exemestane (Aromasin), vorozole (Rivizor), formestane (Lentaron) and fadrozole (Afema). In some embodiments, the aromatase inhibitor used in combination with a diastereomeric compound or salt thereof detailed herein is anastrozole.

In some embodiments, the method of treating estrogen deficiency comprises administering a diastereomeric compound of formula (I) (eg, α-anordrine) or a salt thereof in combination with at least one aromatase inhibitor. . In some embodiments, the method of preventing or alleviating estrogen deficiency symptoms comprises administering a diastereomeric compound (eg, α-anordrine) or salt thereof in combination with at least one aromatase inhibitor.

In some embodiments, a method of preventing or reducing osteoporosis in an individual is provided comprising administering to said individual an effective amount of a diastereomeric compound of formula (I) or salt thereof.

In some embodiments, a method of preventing or reducing osteoporosis in an individual, said method comprising administering to said individual a) an effective amount of a diastereomeric compound of formula (I) or salt thereof; and b) at least one A method is provided comprising administering an effective amount of two additional agents, wherein said additional agent is raloxifene or a functional equivalent thereof. In some embodiments, the additional agent is raloxifene.

In some embodiments, a method of preventing or reducing osteoporosis in an individual, said method comprising administering to said individual a) an effective amount of a diastereomeric compound of formula (I) or salt thereof; and b) at least one A method is provided comprising administering an effective amount of two additional agents, wherein said additional agent is an aromatase inhibitor. In some embodiments, the additional agent is anastrozole.

In some embodiments, the above methods of preventing or reducing osteoporosis further comprise administering to the individual an effective amount of calcium. Suitable amounts of calcium include, but are not limited to, about 0.25 to about 500 mg/day, such as about 10 to about 200 mg/day, about 50 to about 1500 mg/day. In some embodiments, the method further comprises administering an effective amount of vitamin D to the individual. Suitable amounts of vitamin D include, but are not limited to, about 400 to about 800 IU/day, such as about 500 to about 600 IU/day.

In some embodiments, a method of preventing or reducing elevated liver triglycerides in an individual is provided comprising administering to said individual an effective amount of a diastereomeric compound of formula (I) or a salt thereof. be.

In some embodiments, a method of preventing or reducing high liver triglycerides in an individual, said method comprising administering to said individual a) an effective amount of a diastereomeric compound of formula (I) or salt thereof; and b) Methods are provided comprising administering an effective amount of at least one additional agent, wherein said additional agent is raloxifene or a functional equivalent thereof. In some embodiments, the additional agent is raloxifene. In some embodiments, the additional agent is selected from the group consisting of tamoxifen, raloxifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene.

In some embodiments, a method of preventing or alleviating high blood glucose in an individual is provided comprising administering to said individual an effective amount of a diastereomeric compound of formula (I) or a salt thereof. be done.

In some embodiments, a method of preventing or alleviating high blood glucose in an individual, said method comprising administering to said individual a) an effective amount of a diastereomeric compound of formula (I) or a salt thereof; and b. ) administering an effective amount of at least one additional agent, wherein said additional agent is raloxifene or a functional equivalent thereof. In some embodiments, the additional agent is raloxifene. In some embodiments, the additional agent is selected from the group consisting of tamoxifen, raloxifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene.

In some embodiments, a method of preventing or reducing vulvovaginal atrophy in an individual is provided comprising administering to said individual an effective amount of a diastereomeric compound of formula (I) or salt thereof. be.

In some embodiments, a method of preventing or reducing vulvovaginal atrophy in an individual, said method comprising administering to said individual a) an effective amount of a diastereomeric compound of formula (I) or a salt thereof; and b) Methods are provided comprising administering an effective amount of at least one additional agent, wherein said additional agent is raloxifene or a functional equivalent thereof. In some embodiments, the additional agent is raloxifene. In some embodiments, the additional agent is selected from the group consisting of tamoxifen, raloxifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene.

In some embodiments, a method of preventing or reducing vulvovaginal atrophy in an individual, said method comprising administering to said individual a) an effective amount of a diastereomeric compound of formula (I) or a salt thereof; and b) Methods are provided comprising administering an effective amount of at least one additional agent, wherein said additional agent is an aromatase inhibitor. In some embodiments, the additional agent is selected from the group consisting of, but not limited to, anastrozole or a functional equivalent thereof.

In some embodiments, a method of preventing or reducing weight gain in an individual is provided comprising administering to said individual an effective amount of a diastereomeric compound of formula (I) or salt thereof. .

In some embodiments, a method of preventing or reducing weight gain in an individual, said method comprising administering to said individual a) an effective amount of a diastereomeric compound of formula (I) or salt thereof; and b) at least Methods are provided comprising administering an effective amount of one additional agent, wherein said additional agent is raloxifene or a functional equivalent thereof. In some embodiments, the additional agent is raloxifene. In some embodiments, the additional agent is selected from the group consisting of tamoxifen, raloxifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene.

In some embodiments, a method of preventing or reducing high blood triglyceride levels in an individual is provided comprising administering to said individual an effective amount of a diastereomeric compound of formula (I) or a salt thereof. be done.

In some embodiments, a method of preventing or reducing high blood triglycerides in an individual, said method comprising administering to said individual a) an effective amount of a diastereomeric compound of formula (I) or a salt thereof; and b. ) administering an effective amount of at least one additional agent, wherein said additional agent is raloxifene or a functional equivalent thereof. In some embodiments, the additional agent is raloxifene. In some embodiments, the additional agent is selected from the group consisting of tamoxifen, raloxifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene.

である、実質的に純粋なジアステレオマー化合物の有効量を投与することを含む方法が本明細書で提供される。 In certain embodiments, a method of preventing or reducing high liver triglycerides in an individual, wherein said individual comprises (2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A- of formula (Ia) nor-5α-androstane (α-anordrin)

Provided herein are methods comprising administering an effective amount of a substantially pure diastereomeric compound that is

である、実質的に純粋なジアステレオマー化合物の有効量を投与することを含む方法が本明細書で提供される。 In certain embodiments, a method of preventing or reducing high blood triglycerides in an individual, wherein said individual comprises (2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A of Formula (Ia) -nor-5α-androstane (α-anordrin)

Provided herein are methods comprising administering an effective amount of a substantially pure diastereomeric compound that is

である、実質的に純粋なジアステレオマー化合物の有効量を投与することを含む方法が本明細書で提供される。 In certain embodiments, a method of preventing or reducing osteoporosis in an individual, wherein said individual comprises (2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor- of formula (Ia) 5α-androstane (α-anordrin)

Provided herein are methods comprising administering an effective amount of a substantially pure diastereomeric compound that is

である、実質的に純粋なジアステレオマー化合物の有効量を投与することを含む方法が本明細書で提供される。 In certain embodiments, a method of preventing or reducing vulvovaginal atrophy in an individual, comprising administering to said individual (2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A- of formula (Ia) nor-5α-androstane (α-anordrin)

Provided herein are methods comprising administering an effective amount of a substantially pure diastereomeric compound that is

である、実質的に純粋なジアステレオマー化合物の有効量を投与することを含む方法が本明細書で提供される。 In certain embodiments, a method of preventing or alleviating high blood glucose in an individual, comprising administering to said individual (2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A of formula (Ia) -nor-5α-androstane (α-anordrin)

Provided herein are methods comprising administering an effective amount of a substantially pure diastereomeric compound that is

である、実質的に純粋なジアステレオマー化合物の有効量を投与することを含む方法が本明細書で提供される。 In certain embodiments, a method of preventing or reducing weight gain in an individual, wherein said individual comprises (2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor of formula (Ia) -5α-androstane (α-anordrin)

Provided herein are methods comprising administering an effective amount of a substantially pure diastereomeric compound that is

In the above method, the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 98% or greater.

In the above methods, the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 99% or greater, about 99.5% or greater, or about 99.9% or greater.

However, side effects seen with traditional estrogen replacement therapy (ERT), aromatase inhibitors and SERMs remain a concern for some researchers (12). Tamoxifen is marketed as an antagonist of the estrogen classical pathway for treating breast cancer patients and has also been reported as an agonist of ER-α36, potentially causing anti-estrogen therapy resistance, while at the same time reducing endometrial epithelial cell stimulate the growth of uterus, resulting in endometrial cancer (7, 8). Raloxifene was marketed as an upgraded version of tamoxifen with the advantage of fewer side effects and prevention of postmenopausal symptoms such as osteoporosis. However, raloxifene can still cause severe side effects common with tamoxifen treatment, such as thromboembolism and non-alcoholic steatohepatitis (NASH) (9,10). The detailed mechanisms responsible for the side effects caused by either raloxifene or tamoxifen remain unclear. Ipriflavone is a plant hormone derivative whose metabolites bind to ER-αLBD with lower affinity than E2 and exhibit less estrogenic effects. Metabolites of ipriflavone and isoflavone exhibit binding affinity and activity for ER-β and E2, and these metabolites are utilized as medicines to prevent osteoporosis in several countries. At least one clinical trial failed to support their efficacy (11). Aromatase inhibitors have also been launched for ER-positive breast cancer in postmenopausal women. It also inhibited the synthesis of estrogen in bone, resulting in osteoporosis.

A method of reducing the side effects of at least one additional agent in an individual, comprising administering to said individual an effective amount of a diastereomeric compound of formula (I) or a salt thereof when used in combination with said at least one additional agent. Provided herein are methods comprising administering In some embodiments, diastereomeric compounds of formula (I) or salts thereof used in combination with tamoxifen may reduce the side effects of tamoxifen. For example, in the present application, it has been observed that alpha-anordrine used in combination with tamoxifen reduces the side effects of tamoxifen, resulting in an effective combination therapy for the treatment of estrogen deficiency or prevention or relief of estrogen deficiency symptoms. there is In some embodiments, diastereomeric compounds of formula (I) or salts thereof used in combination with at least one aromatase inhibitor may reduce the side effects of the aromatase inhibitor.

である、実質的に純粋なジアステレオマー化合物の有効量を投与することを含む方法が本明細書で提供される。 In certain embodiments, a method of reducing side effects of tamoxifen in an individual comprising administering to the individual (2α,17α)-diethynyl-(2β,17β)-diol-dipropionate of formula (Ia) in combination with tamoxifen -A-nor-5α-androstane (α-anordrin)

Provided herein are methods comprising administering an effective amount of a substantially pure diastereomeric compound that is

である、実質的に純粋なジアステレオマー化合物の有効量を投与することを含む方法が本明細書で提供される。 In certain embodiments, a method of reducing the side effects of at least one aromatase inhibitor in an individual, comprising administering to said individual, (2α,17α)-diethynyl-(of formula (Ia) in combination with said aromatase inhibitor) 2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin)

Provided herein are methods comprising administering an effective amount of a substantially pure diastereomeric compound that is

In the above method, the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 98% or greater.

In the above methods, the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 99% or greater, about 99.5% or greater, or about 99.9% or greater.
Mode of administration

In the context of combination therapy, a composition comprising a diastereomeric compound of formula (I) or salt thereof and an additional agent may be administered simultaneously (ie co-administered) and/or sequentially (ie sequential administration).

In some embodiments, a diastereomeric compound of Formula (I) or salt thereof and an additional agent (including certain agents described herein) are co-administered. The term "co-administration," as used herein, means that the diastereomeric compound of formula (I) or salt thereof and the additional agent are administered for about 15 minutes or less (e.g., about 10 minutes or less, about 5 minutes or less). or about 1 minute or less). When the drugs are co-administered, the diastereomeric compound of formula (I) or salt thereof and the additional agent are in the same composition (e.g., both the diastereomeric compound of formula (I) or salt thereof and the additional agent are compositions (e.g., pharmaceutical compositions included herein) or separate compositions (e.g., diastereomeric compounds of formula (I) or salts thereof and additional agents are included in separate compositions). obtain.

In some embodiments, the diastereomeric compound of Formula (I) or salt thereof and the additional agent are administered sequentially. The term "sequential administration," as used herein, means that the diastereomeric compound of formula (I) or salt thereof and the additional agent are administered for more than about 15 minutes (e.g., about 20 minutes, about 30 minutes, administered at time intervals greater than about 40 minutes, about 50 minutes, about 60 minutes or any longer. Either the diastereomeric compound of formula (I) or a salt thereof or the additional agent may be administered first. The diastereomeric compound of formula (I) or salt thereof and the additional agent may be contained in separate compositions which may be in the same or different packaging.

In some embodiments, the administration of the diastereomeric compound of formula (I) or salt thereof and the additional agent is simultaneous, i.e., the period of administration of the diastereomeric compound of formula (I) or salt thereof and the additional agent is overlap each other. In some embodiments, the diastereomeric compound of Formula (I) or salt thereof is administered for at least one cycle (eg, any of at least two, three, or four cycles) prior to administration of the additional agent. In some embodiments, the additional agent is administered for at least any of 1, 2, 3, or 4 weeks. In some embodiments, the administration of the diastereomeric compound of formula (I) or salt thereof and the additional agent are administered at about the same time (e.g., on any one of days 1, 2, 3, 4, 5, 6 or 7). within the period). In some embodiments, the administration of the diastereomeric compound of formula (I) or salt thereof and the additional agent are administered at about the same time (e.g., on any one of days 1, 2, 3, 4, 5, 6 or 7). within the period). In some embodiments, the administration of the additional agent(s) is after completion of administration of the diastereomeric compound of formula (I) or salt thereof (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9 , for any one period of 10, 11 or 12 months). In some embodiments, administration of the additional agent is initiated after administration of the diastereomeric compound of formula (I) or salt thereof (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9 , 10, 11 months or we months). In some embodiments, administration of the diastereomeric compound of Formula (I) or salt thereof and the additional agent begin and end about the same time. In some embodiments, administration of the diastereomeric compound of formula (I) or salt thereof and the additional agent are initiated at about the same time, and administration of the additional agent is initiated at about the same time as administration of the diastereomeric compound of formula (I) or salt thereof. Continued after termination (eg, for any one period of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months). In some embodiments, administration of the diastereomeric compound of formula (I) or salt thereof and the additional agent are stopped at about the same time, and administration of the additional agent is stopped at about the same time as administration of the diastereomeric compound of formula (I) or salt thereof. After initiation (eg, after any one period of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 months or we months).

The frequency of administration of the diastereomeric compounds of formula (I) or salts thereof and/or additional agents may be adjusted during the course of treatment based on the judgment of the administering physician. When administered separately, the diastereomeric compound of formula (I) or salt thereof and the additional agent may be administered at different dosing frequencies or intervals. For example, a diastereomeric compound of formula (I) or salt thereof may be administered weekly, while the additional agent may be administered more or less frequently. Various formulations and devices for achieving sustained release are known in the art. Exemplary dosing frequencies are provided further herein.

A diastereomeric compound of formula (I) or a salt thereof and the additional agent may be administered using the same route of administration or different routes of administration. Exemplary routes of administration are provided further herein. In some embodiments (for both simultaneous and sequential administration) the diastereomeric compound of formula (I) or salt thereof and the additional agent are administered in a predetermined ratio. For example, in some embodiments, the weight ratio of the diastereomeric compound of Formula (I) or salt thereof and the additional agent is about 1:1. In some embodiments, the weight ratio can be from about 0.001 to about 1 to about 1000 to about 1 or from about 0.01 to about 1 to about 100 to about 1. In some embodiments, the weight ratio of the diastereomeric compound of Formula (I) or salt thereof and the additional agent is about 100:1, about 50:1, about 30:1, about 10:1, about 9: less than any of 1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1 and about 1:1. In some embodiments, the weight ratio of the diastereomeric compound of Formula (I) or salt thereof and the additional agent is about 1:1, about 2:1, about 3:1, about 4:1, about 5: greater than any of 1, about 6:1, about 7:1, about 8:1, about 9:1, about 30:1, about 50:1, about 100:1. Other ratios are contemplated.

The required dose of the diastereomeric compound of formula (I) or salt thereof and/or the additional agent may (but is not necessarily) lower than the dose normally required when each agent is administered alone. do not have). Thus, in some embodiments, a subtherapeutic amount of a diastereomeric compound of formula (I) or salt thereof and/or additional agent is administered. A "sub-therapeutic amount" or "sub-therapeutic level" is less than the therapeutic amount (i.e., the amount normally used when the diastereomeric compound of formula (I) or salt thereof and/or the additional agent is administered alone). lesser). A reduction can be reflected in terms of the amount administered at a given dose and/or the amount administered over a given period of time (decreased frequency).

In some embodiments, at least about 5%, about 10%, about 20%, about 30% of the normal dose of the diastereomeric compound of formula (I) or salt thereof required to produce the same degree of treatment; Sufficient additional agent is administered to enable a percentage reduction of either about 50%, about 60%, about 70%, about 80%, about 90% or more. In some embodiments, at least about 5%, about 10%, about 20%, about 30%, about 50%, about 60%, about 70% of the normal dose of the additional agent required to produce the same degree of treatment Sufficient diastereomeric compounds of formula (I) or salts thereof are administered to enable a percentage reduction of either %, about 80%, about 90% or more.

In some embodiments, the doses of both the diastereomeric compound of formula (I) or salt thereof and the additional agent are reduced compared to the corresponding normal doses of each when administered alone. In some embodiments, both the diastereomeric compound of Formula (I) or salt thereof and the additional agent are administered at sub-therapeutic amounts (ie, reduced levels). In some embodiments, the dose of the diastereomeric compound of formula (I) or salt thereof and/or the additional agent is substantially below the established maximum toxic dose (MTD). For example, the dose of the diastereomeric compound of formula (I) or salt thereof and/or the additional agent is less than about 50%, less than about 40%, less than about 30%, less than about 20% or less than about 10% of the MTD. be.

In some embodiments, the dose of the diastereomeric compound of formula (I) or salt thereof and/or the dose of the additional agent is higher than the dose normally required when each agent is administered alone. For example, in some embodiments, the dose of the diastereomeric compound of formula (I) or salt thereof and/or the additional agent is substantially higher than the established maximum toxic dose (MTD). For example, the dose of the diastereomeric compound of formula (I) or salt thereof and/or the additional agent is about 50%, about 40% of the MTD of the agent when administered alone
, greater than about 30%, about 20%, or about 10%.

In some embodiments, the amount of diastereomeric compound of formula (I) or salt thereof (alone or in combination with additional agents) falls within any of the following ranges: about 0.1 to about 0. .5 mg, about 0.5 to about 5 mg, about 5 to about 10 mg, about 10 to about 15 mg, about 15 to about 20 mg, about 20 to about 25 mg, about 20 to about 50 mg, about 25 to about 50 mg, about 50- about 75 mg, about 50 to about 100 mg, about 75 to about 100 mg, about 100 to about 125 mg, about 125 to about 150 mg, about 150 to about 175 mg, about 175 to about 200 mg, about 200 to about 225 mg, about 225 to about 250 mg , about 250 to about 300 mg, about 300 to about 350 mg, about 350 to about 400 mg, about 400 to about 450 mg, or about 450 to about 500 mg. In some embodiments, the amount of diastereomeric compound of formula (I) or salt thereof (eg, in unit dosage form) ranges from about 5 mg to about 500 mg, such as from about 30 mg to about 300 mg or from about 50 mg to about 200 mg. is.

In some embodiments, the amount of diastereomeric compounds of formula (I) or salts thereof (alone or in combination with additional agents) is at least about 0.01 mg/kg, about 0.05 mg/kg, about 0 .1 mg/kg, about 0.25 mg/kg, about 0.5 mg/kg, about 1 mg/kg, about 2.5 mg/kg, about 3.5 mg/kg, about 5 mg/kg, about 6.5 mg/kg, including either about 7.5 mg/kg, about 10 mg/kg, about 15 mg/kg or about 20 mg/kg. In some embodiments, the amount of diastereomeric compound of formula (I) or salt thereof (alone or in combination with additional agents) is at least about 0.01 mg/kg/day, about 0.05 mg/kg/day about 0.1 mg/kg/day, about 0.25 mg/kg/day, about 0.5 mg/kg/day, about 1 mg/kg/day, about 2.5 mg/kg/day, about 3.5 mg/day kg/day, about 5 mg/kg/day, about 6.5 mg/kg/day, about 7.5 mg/kg/day, about 10 mg/kg/day, about 15 mg/kg/day, or about 20 mg/kg/day including any

In some embodiments, the amount of additional agent is at least about 0.01 mg/kg, about 0.05 mg/kg, about 0.1 mg/kg, about 0.25 mg/kg, about 0.5 mg/kg, about 1 mg/kg, about 2.5 mg/kg, about 3.5 mg/kg, about 5 mg/kg, about 6.5 mg/kg, about 7.5 mg/kg, about 10 mg/kg, about 15 mg/kg or about 20 mg/kg kg. In some embodiments, the amount of diastereomeric compound of formula (I) or salt thereof (alone or in combination with additional agents) is at least about 0.01 mg/kg/day, about 0.05 mg/kg/day about 0.1 mg/kg/day, about 0.25 mg/kg/day, about 0.5 mg/kg/day, about 1 mg/kg/day, about 2.5 mg/kg/day, about 3.5 mg/day kg/day, about 5 mg/kg/day, about 6.5 mg/kg/day, about 7.5 mg/kg/day, about 10 mg/kg/day, about 15 mg/kg/day, or about 20 mg/kg/day including any

Exemplary dosing frequencies of diastereomeric compounds of formula (I) or salts thereof (and additional agents) include, but are not limited to, once every two weeks, once every three weeks, once every four weeks. , once every 6 weeks or once every 8 weeks. In some embodiments, the composition is administered at least about once, about twice, about three times, about four times, about five times, about six times, or about seven times a week (i.e., daily) about It is administered either three times or about twice daily. In some embodiments, the interval between each administration is about 6 months, about 3 months, about 1 month, about 20 days, about 15 days, about 12 days, about 10 days, about 9 days, about less than any of 8 days, about 7 days, about 6 days, about 5 days, about 4 days, about 3 days, about 2 days or about 1 day. In some embodiments, the interval between each administration is about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 8 months, or about Any longer than 12 months. In some embodiments, there are no rest periods in the dosing schedule. In some embodiments, the interval between each administration is about 1 week or less.

Administration of the diastereomeric compound of formula (I) or salt thereof (and additional agents) can be extended for extended periods of time, eg, from about 1 month up to about 7 years. In some embodiments, the composition has at least about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 18, about 24 , about 30, about 36, about 48, about 60, about 72 or about 84 months.

In some embodiments, the individual has at least about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10 times are treated with any of the treatment cycles of

The frequency of administration of the additional agent may be the same as or different from the frequency of administration of the diastereomeric compound of formula (I) or salt thereof. Exemplary frequencies are provided above.

The diastereomeric compounds of formula (I) or salts thereof (and further agents) described herein can be administered, for example, orally, intravenously, intraarterially, intraperitoneally, intrapulmonarily, by inhalation, intravesicular, intramuscularly. , intratracheal, subcutaneous, intraocular, intrathecal, transmucosal, and transdermal. In some embodiments, sustained release formulations of the composition may be used.

Combinations of dosing configurations described herein may be used. The combination therapies described herein can be performed alone or in conjunction with other treatments such as surgery, irradiation, chemotherapy, immunotherapy and gene therapy. Additionally, those at higher risk of developing a proliferative disease may receive treatment to prevent and/or delay the onset of the disease.

As will be appreciated by those skilled in the art, appropriate doses of additional agents approximate those already used in clinical therapy where the additional agents are administered alone or in combination with additional agents. Variation in dosage may occur depending on the condition being treated. As noted above, in some embodiments the additional agent may be administered at reduced levels.
kits and medicines

The invention also provides medicaments, kits and unit dosages useful for the methods described herein. Any use described herein is also provided, regardless of the context of use as a pharmaceutical and/or use for the manufacture of a medicament.

In some embodiments, compositions may be in unit dosage form (such as an oral unit dosage form). Suitable unit dosage forms include, but are not limited to, capsules, tablets, pills, caplets, gels, liquids (eg suspensions, solutions, emulsions), powders or other particulate matter.

In some embodiments, the weight ratio of the diastereomeric compound of formula (I) or salt thereof and the at least one additional agent in the composition is about 1:1. In some embodiments, the weight ratio is about 0.001 to about 1 to about 1000 to about 1 or about 0.01 to about 1 to about 100 to about 1. In some embodiments, the weight ratio of the diastereomeric compound of Formula (I) or salt thereof and the additional agent is about 100:1, about 50:1, about 30:1, about 10:1, about 9: less than any of 1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1 and about 1:1. In some embodiments, the weight ratio of the diastereomeric compound of Formula (I) or salt thereof and the additional agent is about 1:1, about 2:1, about 3:1, about 4:1, about 5: greater than any of 1, about 6:1, about 7:1, about 8:1, about 9:1, about 30:1, about 50:1, about 100:1. In some embodiments, the weight ratio of the diastereomeric compound of formula (I) or salt thereof to the additional agent in the composition is from about 1:20 to about 20:1 (eg, from about 10:1 to about 1:10 or from about 1:10 to about 1:15).

In another aspect, kits are provided comprising a diastereomeric compound of formula (I) or a salt thereof and an additional agent, either in separate containers or in the same container. Kits of the invention comprise one or more containers containing a diastereomeric compound of formula (I) or a salt thereof (or unit dosage form and/or product) and/or at least one additional agent; In some embodiments, further comprising instructions for use according to any of the methods described herein. The kit may further include instructions for selecting an appropriate individual for treatment. Instructions supplied with kits of the invention are typically on a label or package insert (e.g., paper included in the kit), but machine readable instructions (e.g., magnetic storage disk or optical Instructions on a storage disk) are also acceptable.

In some embodiments, the kit is selected from the group consisting of a) an effective amount of a diastereomeric compound of formula (I) or a salt thereof and b) tamoxifen, raloxifene or functional equivalents thereof and an aromatase inhibitor. and an effective amount of at least one additional agent. In some embodiments, the kit is selected from the group consisting of a) an effective amount of a diastereomeric compound of formula (I) or a salt thereof and b) tamoxifen, raloxifene or functional equivalents thereof and an aromatase inhibitor. and c) a diastereomeric compound of formula (I) or a salt thereof and a further agent for the treatment of cancer (or other uses described herein). Also included are instructions for sequential or co-administration.

The diastereomeric compound of formula (I) or salt thereof and the additional agent may be in separate containers or in a single container. The kit may contain one separate composition, or two or more, one composition containing the diastereomeric compound of formula (I) or a salt thereof and one composition containing the additional agent It is understood that it can include the composition of

Kits of the invention are suitably packaged. Suitable packaging includes, but is not limited to, vials, bottles, jars and flexible packaging such as sealed Mylar or plastic bags. Kits may optionally provide additional components such as buffers and interpretive information. Accordingly, the present application also provides products including vials (eg, sealed vials), bottles, jars, flexible packaging, and the like.

Instructions for use of the diastereomeric compounds of formula (I) or salts thereof generally include information regarding dosages, dosing schedules and routes of administration for the desired treatment. The containers may be unit doses, bulk packages (eg, multi-dose packages) or sub-unit doses. For example, for an extended period of time (e.g., 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months or Kits containing sufficient dosages of the diastereomeric compounds of formula (I) or salts thereof disclosed herein to provide effective treatment of an individual for a period of time exceeding The kit may also include multiple unit doses of the diastereomeric compound of formula (I) or salts thereof, a pharmaceutical composition, and instructions for use, for storage and use in pharmacies, such as hospital pharmacies and compounding pharmacies. It can be packaged in sufficient quantity.

Those skilled in the art will recognize that several embodiments are possible within the scope and spirit of the invention. The invention will now be described in more detail with reference to the following non-limiting examples. The following examples further illustrate the invention, but should, of course, not be construed as limiting its scope in any way.

Although the invention has been described and illustrated in some detail, the disclosure has been made by way of example only and any combination of parts may be incorporated without departing from the spirit and scope of the invention as defined in the appended claims. It is understood that many variations in and assortment may be made.
Example 1: Synthesis example

The chemical synthesis of α-anoldrin can be carried out according to Method I below (shown in Scheme I), which is incorporated herein, or by US Pat. No. 5,001,120 and Organic Letters, 2007, 9(9); 1643 It can be prepared as described using the steps detailed in Method II (Scheme II) by reference to publication No.-6.

Reaction Step I: Preparation of Jones Reagent: 57 g CrO ₃ was dissolved in 400 ml H ₂ O, then 85 ml H ₂ SO ₄ was slowly added. The resulting Jones reagent was then cooled to 40°C. 50 g of 5α-androstane and 200 ml of acetic acid were added to a 1 L three-necked flask. The mixture was warmed to 55-60° C., stirred and dissolved under a nitrogen atmosphere. Jones reagent was added dropwise within 1 hour. The reaction temperature was raised to 90° C., reacted for 1 hour, and acetic acid was distilled under reduced pressure. The residue was filtered, washed with H ₂ O and dried to give 50 g of diacid product (b).

Reaction Step I: Preparation of Jones Reagent: 57 g CrO ₃ was dissolved in 400 ml H ₂ O, then 85 ml H ₂ SO ₄ was slowly added. The resulting Jones reagent was then cooled to 40°C. 50 g of 5α-androstane and 200 ml of acetic acid were added to a 1 L three-necked flask. The mixture was warmed to 55-60° C., stirred and dissolved under a nitrogen atmosphere. Jones reagent was added dropwise within 1 hour. The reaction temperature was raised to 90° C., reacted for 1 hour, and acetic acid was distilled under reduced pressure. The residue was filtered, washed with H ₂ O and dried to give 50 g of diacid product (b).

Reaction Step II: 30 g of diacid product (b) and 600 mL of acetic anhydride were dissolved in a 1 L three-necked flask, added to 24 g of sodium acetate, stirred and refluxed for 4 hours. The residue was filtered and dried to give 18 g of diketone product (c).

Reaction Steps III-1 and IV-1: Preparation of Potassium Acetylene Reagent: 15 g of KOH solution (20% w/w) was cooled to 4° C. in a 250 mL 3-necked flask and acetylene was injected to a stable mass. . 6 g of diketone product (c) was dissolved in 42 mL of THF and then added to the 3-necked flask at 25° C. over 2 hours. The product was diluted with _H2O and acidified with HCl. The residue was filtered, washed with H ₂ O and dried to give 5.9 g of 2,17α-diethynyl-A-nor-5a-androstane-2,17β-diol product (e).

Reaction step V: 2.8 g 2,17α-diethynyl-A-nor-5a-androstane-2,17β diol product (e), 4.5 mL propionic acid and 7.8 mL propionic anhydride to 100 mL Mix in flask and then stir for 3 hours. 30 mL of H ₂ O and 40 mL of ethyl acetate were added to the flask, stirred, and the water and organic phases were separated. The organic phase was washed 5 times with water and 10 mL of n-hexane was added. The residue was filtered and dried to give 2.0 g mixture of products α- and β-anordrin (fα and fβ respectively).

According to some embodiments of the present invention, the asymmetric synthesis of α-anoldrin is described using the steps detailed in Method II below (as shown in Scheme II). can be prepared as

Similar to Method I, reaction steps I and II above were used to prepare the diketone product (c).

Reaction Step III-2: Add 6 mL of dry THF, 9.2 mL of trimethylsilylacetylene (TMS) to a dry 250 mL three-necked flask, then cool to −40 to −80° C. and add 20 mL of n-buLi to the solution. Add and stir for 30 minutes. 7 mL of tetramethylethylenediamine (TEMED), 6 g of the diketone product (c) from Reaction Step II of Method I, were dissolved in 42 mL of THF and then cooled to -40 to -80°C. The two solutions are mixed together and allowed to react overnight. The reaction was quenched with _NH4Cl solution. 100 ml of ethyl acetate was added to the flask and stirred for 5 minutes. The organic phase was dried to give 4.8 g of product (d-2).

Reaction step IV-2: 4.8 g of product (d-2) was dissolved in a mixture of 15 ml of THF and 17 ml of tetrabutylammonium fluoride (TBAF) solution and allowed to react for 3 hours. THF was distilled. 2α,17α-diethynyl-A-nor-5a-androstane-2β,17β-diol product α-anoldiol (e-2) was dissolved in ethyl acetate. The organic solution was washed three times with distilled water. After removing the ethyl acetate, 3.5 g of 2α,17α-diethynyl-A-nor-5a-androstane-2β,17β-diol product α-anoldiol (e-2) was obtained.

Reaction Step V: 2.8 g of 2α,17α-diethynyl-A-nor-5a-androstane-2β,17β-diol product (e-2), 4.5 mL of propionic acid and 7.8 mL of propionic anhydride The materials were mixed in a 100 mL flask and then stirred for 3 hours. 30 mL of H ₂ O and 40 mL of ethyl acetate were added to the flask, stirred, and the water and organic phases were separated. The organic phase was washed five times with distilled water and 10 mL of n-hexane was added. The residue was filtered and dried to give 2.0 g of a mixture of products α-anordrine (fα).

Silica gel chromatographic analysis indicated that only one diastereomeric compound was synthesized using Method II (FIG. 1B). In contrast, using Method I, a mixture of diastereomeric compounds was synthesized (Fig. 1A).

NMR Analysis: Fraction 1 (F1) and Fraction 2 (F2) are collected and dissolved in CCl ₄ (50 mg/mL) for NMR analysis. Figures 2A and 2B showed α-anordrin from fraction 1 (F1) and β-anordrin from fraction 2 (F2), respectively.
Example 2: Effect of α-Anordrin or β-Anordrin

In this example, studies demonstrate the estrogenic effects of α- and β-anordrine on estrogen-modulated metabolic signaling and tissue atrophy. Ovaries of 7-week-old mice were surgically excised. One week after surgery, mice were fed with isoflavones, α-anordrin or β-anordrin. Body weight, blood glucose and food intake were measured monthly. After 3 months the mice were sacrificed. Legs and vertebrae, liver, uterus and vagina were harvested.

Wet weight and H&E staining of the uterus and vagina showed that α-anordrine, but not β-anordrine, prevented vulvovaginal atrophy (Fig. 4). α-Anoldrin-treated OVX mice showed weaker symptoms of intrauterine atrophy compared to mice treated with β-anoldrin and the blank group (FIG. 3). α-Anoldrin showed lower blood glucose, TG and body weight (FIGS. 5A and 5B), respectively, whereas initial body weight and food intake (FIGS. 5C and 5D), respectively, were significantly lower than β-anoldrin. there was no difference. These data indicated that the differences were due to changes in energy consumption. Liver H&E sections and statistical analysis showed that α-anordrine, but not β-anordrine, decreased liver TG content in OVX mice (FIG. 6). MicroCT results show that α-anordrin, but not β-anordrin, prevented osteoporosis in OVX mice (Fig. 7).

Tamoxifen was the first FDA-approved drug for breast cancer patients with positive-expressing ER. However, tamoxifen also induces side effects such as endometrial cancer and NASH. Importantly, for women with ER-positive (ER ⁺ ) cancer, continuation of tamoxifen treatment for up to 10 years rather than discontinuing it at 5 years was associated with further reductions in recurrence and mortality, especially after 10 years. Bring. The inventors have found that anoldrine can abrogate tamoxifen-induced endometrial epithelial cell (EEC) mitosis. We further tested whether α- or β-anordrin abrogates tamoxifen-induced EEC mitosis. Isoflavone (blank), α-anordrin (a-ANO), β-anordrin (b-ANO), α-anordrin + tamoxifen (TAM + a-ANO), β-anordrine + tamoxifen (TAM + b-ANO) or tamoxifen (TAM) The containing food was fed to 7-week-old mice for 6 months. Mice were then sacrificed and the uterus and liver were collected. Uterine and liver tissues were fixed.

By H&E-stained sections, EEC in the TAM+b-ANO and TAM groups
Compared to smooth monolayer EECs in the +a-ANO and sham groups, they were rough and mitotic (Fig. 8A), and fatty deposits in hepatocytes of the TAM +b-ANO and TAM groups were higher than those of the TAM + a-ANO and sham groups. were shown to be higher than those of hepatocytes (Fig. 9A). Statistical analysis of H&E-stained sections showed that α-anordrine, but not β-anordrine, inhibited tamoxifen-induced EEC mitosis (FIG. 8B) and NASH (FIG. 9B).

An aromatase inhibitor was administered to ER-positive breasts in postmenopausal women. It also induced side effects such as tissue atrophy and osteoporosis. Anastrozole is an aromatase inhibitor. It was administered to ER-positive breast cancer in postmenopausal women. Anastrozole (ANA) also induced vaginal atrophy and osteoporosis. Vulvovaginal atrophy usually results in infection of the vagina and urinary tract. As an example, we tested whether α- or β-anordrin abrogates anastrozole-induced uterine and vulvovaginal atrophy and osteoporosis. Isoflavones (blank), α-anordrin (a-ANO), β-anordrin (b-ANO), α-anordrin + anastrozole (ANA + a-ANO), β-anordrin + anastrozole (ANA + b-ANO) or Diet containing strozol (ANA) was fed to 7-week-old mice for 6 months. Mice were then sacrificed and the vagina, legs and vertebrae collected. Tissues were fixed with paraformaldehyde. MicroCT results showed that α-anordrin, but not β-anordrin, suppressed anastrozole-induced osteoporosis (FIGS. 10A, 10B and 10C). Statistical analysis of vaginal wet weight and vulvovaginal H&E-stained sections showed that α-anordrine, but not β-anordrine, suppressed anastrozole-induced vaginal atrophy (FIGS. 11A, 11B and 11C). .
Example 3: Materials and Material Preparation Methods

Tamoxifen and Anastrozole (MPG USP grade) were purchased from Okahata (Shanghai) Trading Co. , Ltd. Epiandrostenestone was purchased from Jiangsu Jiaerke Pharmaceuticals Group, LTD. All other compounds were purchased from Sigma or Aladdin. Mouse food was obtained from Trophic Animal Feed High-tech Co. , Ltd, Nantong, China. Mice were obtained from BK animal Inc. purchased from. Animal experiments were performed at the Southern Center of Pesticide Research, Shanghai.

Glucose Concentration Assay: Glucose concentration in culture medium or whole blood from mouse tail was measured using a glucose assay kit according to the manufacturer's instructions (Yicheng, Beijing).

Ovariectomy (OVX) mouse model construction and drug administration: Ovaries were surgically removed from 6-week-old mice. Three days after surgery, drugs were administered by daily gavage or mixed with food.

Preparation of paraffin sections and HE staining: Mouse tissues were surgically excised and fixed using 4% paraformaldehyde (Beijing Solar Bioscience & Technology co., Ltd.) in 1×DPBS. Paraffin section preparation and HE staining were performed by BK animal model, Inc.; was conducted by the GLP Institute of

Measurement of TG in mouse liver: 30-50 mg of mouse liver was surgically resected and 1
It was homogenized in ml chloroform:methanol (2:1) mixture and extracted using 0.5 ml _ddH2O . The organic phase was transferred to a new tube and air dried. The amount of TG was measured using a kit according to the manufacturer's instructions. An internal standard control was used to correct for errors.

Measurement of EEC Height, Uterine Diameter and Circular Muscle Thickness: EEC height was measured from 20-200× EEC images of paraffin-embedded H&E mouse uterine sections.

Measurement of vulvovaginal wall thickness: The vagina was cut in the center. For preparation of H&E sections, both uterine and vulvovaginal sites were embedded in paraffin in the same orientation. Vulvovaginal wall thickness was measured from x40 magnification images of paraffin-embedded H&E mouse vaginal sections.

Bone Density Assay Using MicroCT: Femurs were fixed in 1×DPBS containing 3% formaldehyde for 2 weeks. After 1 week, the fixative was changed. Density of femurs was measured by Siemens Inveon microCT. HU2000 values were analyzed using Inveon Research Workplace (IRW) under the following measurement conditions: 80 KVP, 500 mA, exposure time 1500 ms; CCD readout settings: axial 2048, binning 2048; FOV transaxial: 19.03 mm, Axial: 19.03 mm, pixel size: 9.29 μm.

Statistical analysis: Tables and figures expressed results as mean + STDEV. Asterisks indicate statistically significant differences calculated using the two-tailed Student's t-test.
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According to preferred embodiments of the present invention, for example, the following are provided.
(Section 1)
Formula (I):

A method for stereospecifically preparing a substantially pure diastereomeric compound of or a salt thereof, wherein
R. ₁ is —OH, —OC(O)—R ^1a or -OC(O)R ^1b COOH and R ₄ is —OH, —OC(O)—R ^4a or -OC(O)R ^4b COOH, where R ^1a and R ^4a are independently hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ cycloalkyl and R ^1b and R ^4b independently -C ₁ -C ₆ alkyl- or -C ₂ -C ₆ is alkenyl;
R. ₂ and R ₅ is -C≡CH;
R. ₆ and R ₇ independently, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ is cycloalkyl;
R. ₈ and R ₉ is independently hydrogen, —OH, —NH ₂ , -NO ₃ , Halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₄ aryl, 5- to 6-membered heteroaryl or 3- to 6-membered heterocyclyl, the method comprising (a) an organometallic reagent R ₁₀ -M, where M is Li, Na or K and R ₁₀ is —OH, halogen or C ₁ -C ₆ C optionally substituted with alkyl ₁ -C ₂₀ is an alkyl),
Formula (II)

The diketone compound of formula (III)

of silylacetylene (where R ^a , R ^b and R ^c is hydrogen, -OH, -OH, halogen or C ₁ -C ₆ C optionally substituted with alkyl ₁ -C ₂₀ alkyl, —OH, halogen or C ₁ -C ₆ C optionally substituted with alkyl ₁ -C ₆ Alkoxy, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₄ independently selected from the group consisting of aryl, 5-6 membered heteroaryl or 3-6 membered heterocyclyl).
(Section 2)
R. ₁ is, -OH, -OC (O) -R ^1a or -OC(O)R ^1b COOH and R ₄ is, -OH, -OC (O) -R ^4a or -OC(O)R ^4b COOH, where R ^1a and R ^4a are independently hydrogen, C ₁ -C ₆ alkyl or C ₂ -C ₆ alkenyl and R ^1b and R ^4b independently, -C ₁ -C ₆ alkyl- or -C ₂ -C ₆ is alkenyl;
R. ₆ and R ₇ independently, C ₁ -C ₆ alkyl or C ₂ -C ₆ is alkenyl;
R. ₈ and R ₉ is independently hydrogen, —OH, —NH ₂ , -NO ₃ , C ₁ -C ₆ alkyl or C ₂ -C ₆ 2. The method according to item 1 above, which is alkenyl.
(Section 3)
R. ₁ is-OC(O)-R ^1a and R ₄ is-OC(O)-R ^4a and R ^1a and R ^4a is both ethyl.
(Section 4)
R. ₁ and R ₄ are both —OH.
(Section 5)
R. ₁ is-OC(O)R ^1b COOH and R ₄ is-OC(O)R ^4b COOH and R ^1b and R ^4b are both -CH ₂ -, -CH ₂ CH ₂ - and -CH=CH-. 3. The method of claim 1 or 2, independently selected from ethyl.
(Section 6)
R. ₆ and R ₇ are both methyl.
(Section 7)
R. ₈ and R ₉ is independently hydrogen or C ₁ -C ₆ 7. The method according to any one of the above items 1 to 6, which is alkyl.
(Section 8)
R. ₈ and R ₉ are both hydrogen.
(Section 9)
R. ^a , R ^b and R ^c independently, C ₁ -C ₆ 9. The method according to any one of the above items 1 to 8, which is alkyl.
(Section 10)
R. ^a , R ^b and R ^c is methyl and said silylacetylene is trimethylsilylacetylene (TMS).
(Item 11)
R. ₁₀ is C ₁ -C ₆ 11. The method according to any one of the above items 1 to 10, which is alkyl.
(Item 12)
R. ₁₀ is n-butyl.
(Item 13)
13. The method according to any one of items 1 to 12, wherein M is Li.
(Item 14)
14. The method of any one of the preceding clauses 1-13, wherein tetramethylethylenediamine (TEMED) is added with the organometallic reagent.
(Item 15)
(b) Formula (IV)

15. The method according to any one of the above items 1 to 14, further comprising removing the silyl group of
(Item 16)
16. The method according to item 15, wherein the silyl group is trimethylsilyl.
(Item 17)
16. The method of claim 15, wherein said removing step is carried out by contact with a deprotecting agent selected from the group consisting of tetrabutylammonium fluoride (TBAF), hydrofluoric acid and potassium fluoride.
(Item 18)
Said substantially pure diastereomeric compound is represented by formula (Ia)

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 19)
19. The method of any one of the preceding clauses 1-18, wherein the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 98% or greater.
(Section 20)
20. Any of the above paragraphs 1-19, wherein the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 99% or greater, about 99.5% or greater, or about 99.9% or greater. or the method described in paragraph 1.
(Section 21)
A method of treatment of estrogen deficiency in an individual, comprising administering to said individual formula (I):

A substantially pure diastereomeric compound of (wherein
R. ₁ is —OH, —OC(O)—R ^1a or -OC(O)R ^1b COOH and R ₄ is —OH, —OC(O)—R ^4a or -OC(O)R ^4b COOH, where R ^1a and R ^4a are independently hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ cycloalkyl and R ^1b and R ^4b independently -C ₁ -C ₆ alkyl- or -C ₂ -C ₆ is alkenyl;
R. ₂ and R ₅ is -C≡CH;
R. ₆ and R ₇ independently, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ is cycloalkyl;
R. ₈ and R ₉ is independently hydrogen, —OH, —NH ₂ , -NO ₃ , Halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₄ aryl, 5-6 membered heteroaryl or 3-6 membered heterocyclyl) or a salt thereof.
(Section 22)
A method of preventing or alleviating estrogen deficiency symptoms in an individual, comprising administering to said individual formula (I):

A substantially pure diastereomeric compound of (wherein
R. ₁ is —OH, —OC(O)—R ^1a or -OC(O)R ^1b COOH and R ₄ is —OH, —OC(O)—R ^4a or -OC(O)R ^4b COOH, where R ^1a and R ^4a are independently hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ cycloalkyl and R ^1b and R ^4b independently -C ₁ -C ₆ alkyl- or -C ₂ -C ₆ is alkenyl;
R. ₂ and R ₅ is -C≡CH;
R. ₆ and R ₇ independently, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ is cycloalkyl;
R. ₈ and R ₉ is independently hydrogen, —OH, —NH ₂ , -NO ₃ , Halogen, C ₁ -C ₆
alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₄ aryl, 5-6 membered heteroaryl or 3-6 membered heterocyclyl) or a salt thereof.
(Section 23)
23. The method of paragraph 22, wherein the estrogen deficiency symptoms are selected from the group consisting of high liver triglycerides, osteoporosis, vulvovaginal atrophy, high blood triglycerides, high blood glucose and weight gain.
(Section 24)
23. The method of paragraph 22, further comprising administering to said individual an effective amount of at least one additional agent, wherein said additional agent is selected from the group consisting of a selective estrogen receptor modulator and an aromatase inhibitor. done, method.
(Section 25)
25. The method of clause 24, wherein said additional agent is tamoxifen.
(Section 26)
25. The method of clause 24, wherein said additional agent is selected from the group consisting of raloxifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene.
(Section 27)
27. The method of any one of paragraphs 24-26, wherein the estrogen deficiency symptoms are selected from the group consisting of high liver triglycerides, osteoporosis, vulvovaginal atrophy, high blood triglycerides, high blood glucose and weight gain.
(Section 28)
28. The method of any one of clauses 24-27, wherein said substantially pure diastereomeric compound of formula (I) or salt thereof and said additional agent are administered sequentially.
(Section 29)
28. The method of any one of clauses 24-27, wherein said substantially pure diastereomeric compound of formula (I) or salt thereof and said additional agent are co-administered.
(Item 30)
A method of reducing the side effects of at least one additional agent in an individual, said method comprising administering to said individual, in combination with said additional agent, Formula (I):

A substantially pure diastereomeric compound of (wherein
R. ₁ is —OH, —OC(O)—R ^1a or -OC(O)R ^1b COOH and R ₄ is —OH, —OC(O)—R ^4a or -OC(O)R ^4b COOH, where R ^1a and R ^4a are independently hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ cycloalkyl and R ^1b and R ^4b independently -C ₁ -C ₆ alkyl- or -C ₂ -C ₆ is alkenyl;
R. ₂ and R ₅ is -C≡CH;
R. ₆ and R ₇ independently, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ ah
lukynyl or C ₃ -C ₆ is cycloalkyl;
R. ₈ and R ₉ is independently hydrogen, —OH, —NH ₂ , -NO ₃ , Halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₄ aryl, 5-6 membered heteroaryl or 3-6 membered heterocyclyl) or salts thereof, wherein said additional agent is selected from the group consisting of selective estrogen receptor modulators and aromatase inhibitors. done, method.
(Item 31)
31. The method of clause 30, wherein said additional agent is tamoxifen.
(Item 32)
32. The method of any one of paragraphs 21-31, wherein said individual is a human.
(Item 33)
R. ₁ is, -OH, -OC (O) -R ^1a or -OC(O)R ^1b COOH and R ₄ is, -OH, -OC (O) -R ^4a or -OC(O)R ^4b COOH, where R ^1a and R ^4a are independently hydrogen, C ₁ -C ₆ alkyl or C ₂ -C ₆ alkenyl and R ^1b and R ^4b independently, -C ₁ -C ₆ alkyl- or -C ₂ -C ₆ is alkenyl;
R. ₆ and R ₇ independently, C ₁ -C ₆ alkyl or C ₂ -C ₆ is alkenyl;
R. ₈ and R ₉ is independently hydrogen, —OH, —NH ₂ , -NO ₃ , C ₁ -C ₆ alkyl or C ₂ -C ₆ 33. The method according to any one of the above items 21 to 32, which is alkenyl.
(Item 34)
R. ₁ is-OC(O)-R ^1a and R ₄ is-OC(O)-R ^4a and R ^1a and R ^4a are both ethyl.
(Item 35)
R. ₁ and R ₄ are both —OH.
(Item 36)
R. ₁ is-OC(O)R ^1b COOH and R ₄ is-OC(O)R ^4b COOH and R ^1b and R ^4b are both -CH ₂ -, -CH ₂ CH ₂ 33. The method of any one of the preceding clauses 21-32 independently selected from - and -CH=CH-.
(Item 37)
R. ₆ and R ₇ are both methyl.
(Item 38)
R. ₈ and R ₉ is independently hydrogen or C ₁ -C ₆ 38. The method according to any one of the above items 21 to 37, which is alkyl.
(Item 39)
R. ₈ and R ₉ are both hydrogen.
(Item 40)
Said substantially pure diastereomeric compound is represented by formula (Ia)

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 41)
41. The method of any one of clauses 21-40, wherein the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 98% or greater.
(Item 42)
42. Any of the above paragraphs 21-41, wherein the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 99% or greater, about 99.5% or greater, or about 99.9% or greater. The method according to item 1.
(Item 43)
Formula (I):

A pharmaceutical composition comprising a substantially pure diastereomeric compound of or a salt thereof, wherein
R. ₁ is —OH or —OC(O)—R ^1a and R ₄ is —OH or —OC(O)—R ^4a , where R ^1a and R ^4a are independently hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ is cycloalkyl; ₂ and R ₅ is -C≡CH;
R. ₆ and R ₇ independently, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or C ₃ -C ₆ is cycloalkyl;
R. ₈ and R ₉ is independently hydrogen, —OH, —NH ₂ , -NO ₃ , Halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₄ A pharmaceutical composition which is aryl, 5-6 membered heteroaryl or 3-6 membered heterocyclyl.
(Item 44)
44. The pharmaceutical composition of Clause 43, further comprising at least one additional agent,
The pharmaceutical composition, wherein the additional agent is selected from the group consisting of selective estrogen receptor modulators and aromatase inhibitors.
(Item 45)
45. The pharmaceutical composition of clause 44, wherein said additional agent is tamoxifen.
(Item 46)
45. The pharmaceutical composition of clause 44, wherein said additional agent is selected from the group consisting of raloxifene, lasofoxifene, bazedoxifene, arzoxifene, olmeloxifene, ospemifene and levormeloxifene.
(Item 47)
45. The pharmaceutical composition according to Clause 44, wherein said additional agent is an aromatase inhibitor.
(Item 48)
48. The pharmaceutical composition according to item 47, wherein the aromatase inhibitor is anastrozole.
(Item 49)
48. The pharmaceutical composition according to item 47, wherein the aromatase inhibitor is selected from the group consisting of anastrozole, letrozole, exemestane, vorozole, formestane and fadrozole.
(Section 50)
Clauses 44-49 above, wherein the weight ratio of said substantially pure diastereomeric compound of formula (I) or salt thereof to said additional agent in said composition is from about 20:1 to about 1:20. A pharmaceutical composition according to any one of claims.
(Item 51)
Said substantially pure diastereomeric compound is represented by formula (Ia)

51. The pharmaceutical composition according to any one of the above items 44 to 50, which is (2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin) of thing.
(Item 52)
52. The pharmaceutical composition of any one of clauses 43-51, wherein said substantially pure diastereomeric compound has a diastereomeric excess (de) of about 98% or greater.
(Item 53)
53. Any of the above paragraphs 43-52, wherein the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 99% or greater, about 99.5% or greater, or about 99.9% or greater. The pharmaceutical composition according to item 1.
(Item 54)
said estrogen deficiency symptom is elevated liver triglycerides and said substantially pure diastereomeric compound is of formula (Ia)

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 55)
said estrogen deficiency symptom is elevated blood triglycerides and said substantially pure diastereomeric compound is represented by formula (Ia)

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 56)
said estrogen deficiency condition is osteoporosis and said substantially pure diastereomeric compound is represented by formula (Ia)

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 57)
said estrogen deficiency symptom is vulvovaginal atrophy and said substantially pure diastereomerization
compound of the formula (Ia)

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 58)
said estrogen deficiency symptom is high blood glucose and said substantially pure diastereomeric compound is represented by formula (Ia)

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 59)
said estrogen deficiency symptom is weight gain and said substantially pure diastereomeric compound is represented by formula (Ia)

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 60)
wherein said further agent is tamoxifen and said substantially pure diastereomeric compound is of Formula (Ia) in combination with tamoxifen

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 61)
Formula (Ia), wherein said further agent is an aromatase inhibitor and said substantially pure diastereomeric compound is combined with said aromatase inhibitor

(2α,17α)-diethynyl-(2β,17β)-diol-dipropionate-A-nor-5α-androstane (α-anordrin).
(Item 62)
62. The method of any one of clauses 54-61, wherein said substantially pure diastereomeric compound has a diastereomeric excess (de) of about 98% or greater.
(Item 63)
63. Any of the above paragraphs 54-62, wherein the substantially pure diastereomeric compound has a diastereomeric excess (de) of about 99% or greater, about 99.5% or greater, or about 99.9% or greater. The method according to item 1.

Claims (1)

The invention described in the specification .

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